Device, system, and method for controllably reducing inflammatory mediators in a subject

ABSTRACT

Devices, systems, and methods are provided for controlling an inflammatory response in a subject. Devices, systems, and methods are provided that alter the functional structure of one or more inflammatory mediators in the peripheral blood of the subject. The device or system is useful in a method for treating an inflammatory disease or condition in the subject.

RELATED APPLICATIONS

For purposes of the USPTO extra-statutory requirements, the presentapplication constitutes a continuation of U.S. patent application Ser.No. 12/380,400, entitled DEVICE, SYSTEM, AND METHOD FOR CONTROLLABLYREDUCING INFLAMMATORY MEDIATORS IN A SUBJECT, naming Roderick A. Hyde,Muriel Y. Ishikawa, Edward K.Y. Jung, Robert Langer, Eric C. Leuthardt,Nathan P. Myhrvold, Elizabeth A. Sweeney and Lowell L. Wood, Jr. asinventors, filed 25 Feb. 2009, which is currently co-pending, or is anapplication of which a currently co-pending application is entitled tothe benefit of the filing date.

For purposes of the USPTO extra-statutory requirements, the presentapplication constitutes a continuation-in-part of U.S. patentapplication Ser. No. 12/380,399 , entitled DEVICE, SYSTEM, AND METHODFOR CONTROLLABLY REDUCING INFLAMMATORY MEDIATORS IN A SUBJECT, namingRODERICK A. HYDE, MURIEL Y. ISHIKAWA, EDWARD K.Y. JUNG, ROBERT LANGER,ERIC C. LEUTHARDT, NATHAN P. MYHRVOLD, ELIZABETH A. SWEENEY, AND LOWELLL . WOOD, JR. as inventors, filed 25 Feb. 2009, which is currentlyco-pending, or is an application of which a currently co-pendingapplication is entitled to the benefit of the filing date.

The United States Patent Office (USPTO) has published a notice to theeffect that the USPTO's computer programs require that patent applicantsreference both a serial number and indicate whether an application is acontinuation or continuation-in-part. Stephen G. Kunin, Benefit ofPrior-Filed Application, USPTO Official Gazette Mar. 18, 2003, availableat http://www.uspto.gov/web/offices/com/sol/og/2003/week11/patbene.htm.The present Applicant Entity (hereinafter “Applicant”) has providedabove a specific reference to the application(s) from which priority isbeing claimed as recited by statute. Applicant understands that thestatute is unambiguous in its specific reference language and does notrequire either a serial number or any characterization, such as“continuation” or “continuation-in-part,” for claiming priority to U.S.patent applications. Notwithstanding the foregoing, Applicantunderstands that the USPTO's computer programs have certain data entryrequirements, and hence Applicant is designating the present applicationas a continuation-in-part of its parent applications as set forth above,but expressly points out that such designations are not to be construedin any way as any type of commentary and/or admission as to whether ornot the present application contains any new matter in addition to thematter of its parent application(s).

All subject matter of the Related Applications and of any and allparent, grandparent, great-grandparent, etc. applications of the RelatedApplications is incorporated herein by reference to the extent suchsubject matter is not inconsistent herewith.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is related to and claims the benefit of theearliest available effective filing date(s) from the following listedapplication(s) (the “Related Applications”) (e.g., claims earliestavailable priority dates for other than provisional patent applicationsor claims benefits under 35 USC §119(e) for provisional patentapplications, for any and all parent, grandparent, great-grandparent,etc. applications of the Related Application(s)). All subject matter ofthe Related Applications and of any and all parent, grandparent,great-grandparent, etc. applications of the Related Applications isincorporated herein by reference to the extent such subject matter isnot inconsistent herewith.

SUMMARY

Devices, systems, and methods are disclosed herein for controlling ormodulating an inflammatory response in a subject. Devices, systems, andmethods are disclosed that alter the functional structure or biologicalactivity of one or more inflammatory mediators in the peripheral bloodof the subject. The device or system is useful in a method for treatingan inflammatory disease or condition in the subject. Diseases orconditions related to either acute or chronic inflammatory responseinclude, but are not limited to, systemic inflammatory responsesyndrome, sepsis, septic shock, multiple organ dysfunction syndrome,ischemia reperfusion, hyperreactive airway disease, (e.g., asthma,chronic obstructive pulmonary disease, rhinitis, sinusitis), allergicreaction, anaphylaxis, pulmonary failure, adult respiratory distresssyndrome (ARDS), allograft rejection, graft versus host disease (GVHD),chronic inflammatory disease, psoriatic arthritis, rheumatoid arthritis,chemical or biological agent exposure (due to warfare, accident, oroccupation), infectious disease, malaria, anthrax, viral hepatitis,viral infection, influenza, smallpox, HIV, myocardial ischemia, orautoimmune disease.

A device is disclosed that includes a sensor configured to detect one ormore inflammatory mediators in peripheral blood of a subject, acontroller in communication with the sensor, and a means for modulatinga physiological effect of the one or more inflammatory mediatorsresponsive to the controller, the controller configured to adjust themodulating means to achieve a target value of the detected one or moreinflammatory mediators in the peripheral blood of the subject. In anaspect, the means for modulating a physiological effect of the one ormore inflammatory mediators can include a treatment chamber configuredto receive at least a portion of the peripheral blood through a flowroute, the controller configured to control flow of peripheral bloodthrough the flow route into the treatment chamber, and the treatmentchamber including one or more reactive components configured to alter afunctional structure of the one or more inflammatory mediators in theperipheral blood. The controller can be configured to control access tothe treatment chamber by the peripheral blood, including access to oneor more reactive components in the treatment chamber. The one or morereactive components can include, but are not limited to, one or more ofa denaturing agent, a degradative agent, binding agent, or an energysource.

In a further aspect, the means for modulating a physiological effect ofthe one or more inflammatory mediators can include the controllerconfigured to control interaction between one or more reactivecomponents and the one or more inflammatory mediators, the one or morereactive components configured to alter a functional structure of theone or more inflammatory mediators in the peripheral blood. Thecontroller can be in communication with a reservoir configured torelease the one or more reactive components into the peripheral blood ofthe subject. The one or more reactive components can include, but arenot limited to, one or more of a denaturing agent, a degradative agent,binding agent, or an energy source. The one or more binding agents caninclude, but are not limited to, one or more of antibodies, receptors,or cognates configured to bind to at least one of the one or moreinflammatory mediators. The one or more binding agents can include oneor more of lectin, binding protein, catalytic antibody, catalyticaptamer, protease conjugate, or photoactivatable agent conjugate. Thetarget value can include a desired concentration of the one or moreinflammatory mediators in the peripheral blood. The target value caninclude a desired range of concentrations of the one or moreinflammatory mediators in the peripheral blood. The target value caninclude a desired ratio of concentrations of two or more inflammatorymediators in the peripheral blood. The target value can include adesired ratio of levels of two or more inflammatory mediators in theperipheral blood.

The sensor and the controller can be configured to control levels of theone or more inflammatory mediators to substantially attain the targetvalue. The sensor and the controller can be configured to control levelsof the one or more inflammatory mediators to limit a deviation from thetarget value. The deviation can be determined by a weighted leastsquares fit. The sensor includes, but is not limited to, a biosensor,chemical sensor, physical sensor, or optical sensor. The sensor caninclude one or more of an aptamer, antibody, or receptor. The sensor caninclude one or more of a recognition-based substrate, an aptamer-basedsubstrate, an antibody-based substrate, surface plasmon resonance,genetically-modified cells, or genetically-modified cells withreceptor-linked signaling. The genetically-modified cells can includereceptor-linked signaling by fluorogen-activating proteins.

The sensor can be configured to target the device to a site having anelevated level of the inflammatory mediators. The sensor can beconfigured to detect cytokines, T-lymphocytes, B-lymphocytes, orantibodies. The sensor can be configured to detect body temperature,vital signs, edema, oxygen level, or pathogen/toxin level of thesubject. The sensor can be configured to detect one or more ofanaphylatoxin, cytokine, chemokine, leukotriene, prostaglandin,complement, coagulation factor, or proinflammatory cytokine. The sensorcan be configured to detect one or more of TNF-α, IL-1, IL-1β, IL-6,IL-8, IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a,exotoxin, or endotoxin.

The one or more binding agents can be configured to sequester at leastone of the one or more inflammatory mediators from the blood. The atleast one of the one or more inflammatory mediators can be sequesteredby the one or more binding agents prior to treatment with one or more ofa denaturing agent, a degradative agent, binding agent, or an energysource. The one or more binding agents can include, but are not limitedto, one or more of antibodies, receptors, or cognates configured to bindto at least one of the one or more inflammatory mediators. The one ormore binding agent includes, but is not limited to, at least one oflectin, binding protein, catalytic antibody, catalytic aptamer, proteaseconjugate, or photoactivatable agent conjugate. The matrix includes, butis not limited to, one or more of a specific binding ligand or ahydrophobic surface. The matrix can include, but is not limited to, oneor more of beads, cells, vesicles, filters, hydrogel polymers,microparticles, nanoparticles, adsorbent, or synthetic polymers. Thespecific binding ligand or the hydrophobic surface can include, but isnot limited to, one or more of nucleic acid aptamers, peptide aptamers,molecular imprinting polymer, antibodies or fragments thereof, highaffinity mimetics, synthetic binding molecules, or receptor bindingmolecules. The matrix can include, but is not limited to, one or more ofa lectin, binding protein, receptor, antibody, catalytic antibody,catalytic aptamer, protease conjugate, or photoactivatable agentconjugate.

In an aspect, the one or more reactive components can be configured todecrease an activity of one or more of TNF-α, IL-1, IL-1β, IL-6, IL-8,IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin,or endotoxin. The one or more reactive components can modulate anactivity of an intermediate which modulates the activity of one or moreof TNF-α, IL-1, IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, IFN-γ, LIF, MIF,MIP-1, MCP-1, C3-a, C5-a, exotoxin, or endotoxin. The one or morereactive components can increase an activity of an intermediate whichdecreases the activity of one or more of TNF-α, IL-1, IL-1β, IL-6, IL-8,IL-10, IL-12, LPB, IFN-γLIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin, orendotoxin. The one or more reactive components can decrease an activityof an intermediate which decreases the activity of one or more of TNF-α,IL-1, IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, INF-γLIF, MIF, MIP-1, MCP-1,C3-a, C5-a, exotoxin, or endotoxin. In an aspect, the one or morereactive components can be configured to modulate an activity of one ormore of anaphylatoxins, cytokines, chemokines, leukotrienes,prostaglandins, complement, coagulation factors, or proinflammatorycytokines.

The energy source can include, but is not limited to, acoustic energy orelectronic energy. The energy source can include ultrasound. The energysource can include high-intensity focused ultrasound. The energy sourcecan include, but is not limited to, at least one of microwaveirradiation, gamma irradiation, electron beam irradiation,vibrational/frequency irradiation, or atmospheric pressure glowdischarge. The vibrational/frequency irradiation can include a set ofdiffering energy inputs specifically directed to the one or moreinflammatory mediators, wherein the set of differing energy inputsselectively resonates a plurality of resonant structures in the one ormore inflammatory mediators, and wherein the resonance controllablyalters or reduces the activity of the one or more inflammatory mediatorsin the peripheral blood of the subject. The one or more inflammatorymediators can be modified with a functional group configured to beresponsive to the set of differing energy inputs. The energy source caninclude, but is not limited to, acoustic energy or electronic energy.The one or more denaturing agents can include, but is not limited to, atleast of an acid, base, solvent, cross-linking agent, chaotropic agent,disulfide bond reducer, enzyme, drug, cell, or radical ion. The one ormore degradative agents can include, but is not limited to, at least oneof an enzyme, coenzyme, enzyme complex, catalytic antibody, proteasome,strong acid, strong base, radical, photoactivatable agent, drug, cell,or radical ion. The catalytic antibody can generate a radical ion. Theone or more reactive components can include an activator or an inhibitorof one or more of a myeloid cell, lymphocyte, or precursor thereof. Theone or more reactive components can include an activator or an inhibitorof one or more of a T-lymphocyte, B-lymphocyte, macrophage, monocyte,neutrophil, or NK cell. The one or more reactive components can includea target-specific release compound including one or more cytokines. Thetarget-specific release compound can include DNA aptamer-crosslinkedhydrogel including the one or more cytokines.

In an aspect, the treatment chamber includes a source for producing theone or more reactive components. The source can include at least onereservoir and at least one producer. The at least one producer caninclude at least one encapsulated cell. The at least one encapsulatedcell can produce the one or more reactive components. The at least oneencapsulated cell can be genetically-engineered. The at least oneencapsulated cell can include, but is not limited to, at least one of amammalian cell, bacterial cell, yeast cell, plant cell, insect cell,artificial cell, or enucleated cell. The at least one encapsulated cellcan include, but is not limited to, one or more of a myeloid cell,lymphocyte, or precursor thereof. The at least one encapsulated cell caninclude, but is not limited to, one or more of a T-lymphocyte,B-lymphocyte, macrophage, monocyte, neutrophil, or NK cell. The at leastone producer can include, but is not limited to, a protein, lipidmicelle, liposome, synthetic polymer, or a combination thereof. The atleast one producer can include a catalytic antibody. The catalyticantibody can include a radical ion generator. In an aspect, thetreatment chamber can include one or more reservoirs including one ormore reactive components.

In an aspect, the one or more reactive components can include at leastone modulator of the one or more inflammatory mediators configured toalter the functional structure of the one or more inflammatory mediatorsin the peripheral blood of the subject. The one or more reactivecomponents can include, but is not limited to, at least one of acytokine, cytokine modulator, matrix metalloproteinase (MMP), orextracellular matrix metalloproteinase inducer (EMMPRIN). The at leastone modulator can include an agonist or antagonist of the one or moreinflammatory mediators. The one or more energy source can include anultrasound source to increase cytokine production. The ultrasound sourcecan provide a pulsed radio frequency ultrasound signal.

The one or more inflammatory mediators can include, but is not limitedto, at least one anaphylatoxin, cytokine, chemokine, leukotriene,prostaglandin, complement, coagulation factor, or proinflammatorycytokine. The one or more inflammatory mediators can include, but is notlimited to, one or more of TNF-α, IL-1, L-1β, IL-6, IL-8, IL-10, IL-12,LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin or endotoxin.

In an aspect, the device is intracorporeal. The device can be configuredto be implanted. The device includes, but is not limited to, a stent,bypass implant, nanostructure or microstructure. The device can beconfigured to be implanted relative to an organ or tissue in thesubject. In an aspect, the device is extracorporeal. The extracorporealdevice can include, but is not limited to, a dialysis device,hemoperfusion device, apheresis device, intravenous device, or patchdevice.

A method for treating an inflammatory disease or condition in a subjectis disclosed that includes providing a device configured to communicatewith at least a portion of a peripheral blood of the subject, the deviceincluding a sensor configured to detect one or more inflammatorymediators in peripheral blood of a subject, a controller incommunication with the sensor, a means for modulating a physiologicaleffect of the one or more inflammatory mediators responsive to thecontroller, the controller configured to adjust the modulating means toachieve a target value of the detected one or more inflammatorymediators in the peripheral blood of the subject. In an aspect, themeans for modulating a physiological effect of the one or moreinflammatory mediators can include a treatment chamber configured toreceive at least a portion of the peripheral blood through a flow route,the controller configured to control flow of peripheral blood throughthe flow route into the treatment chamber, and the treatment chamberincluding one or more reactive components configured to alter afunctional structure of the one or more inflammatory mediators in theperipheral blood. The controller can be configured to control access tothe treatment chamber by the peripheral blood, including access to oneor more reactive components in the treatment chamber. The one or morereactive components can include, but are not limited to, one or more ofa denaturing agent, a degradative agent, binding agent, or an energysource

In a further aspect, the means for modulating a physiological effect ofthe one or more inflammatory mediators can include the controllerconfigured to control interaction between one or more reactivecomponents and the one or more inflammatory mediators, the one or morereactive components configured to alter a functional structure of theone or more inflammatory mediators in the peripheral blood. Thecontroller can be in communication with a reservoir configured torelease the one or more reactive components into the peripheral blood ofthe subject. The one or more reactive components can include, but arenot limited to, one or more of a denaturing agent, a degradative agent,binding agent, or an energy source. The one or more binding agents caninclude, but are not limited to, one or more of antibodies, receptors,or cognates configured to bind to at least one of the one or moreinflammatory mediators. The one or more binding agents can include oneor more of lectin, binding protein, catalytic antibody, catalyticaptamer, protease conjugate, or photoactivatable agent conjugate. Thetarget value can include a desired concentration of the one or moreinflammatory mediators in the peripheral blood. The target value caninclude a desired range of concentrations of the one or moreinflammatory mediators in the peripheral blood. The target value caninclude a desired ratio of concentrations of two or more inflammatorymediators in the peripheral blood. The target value can include adesired ratio of levels of two or more inflammatory mediators in theperipheral blood.

The sensor and the controller can be configured to control levels of theone or more inflammatory mediators to substantially attain the targetvalue. The sensor and the controller can be configured to control levelsof the one or more inflammatory mediators to limit a deviation from thetarget value. The deviation can be determined by a weighted leastsquares fit. The sensor includes, but is not limited to, a biosensor,chemical sensor, physical sensor, or optical sensor. The sensor caninclude one or more of an aptamer, antibody, or receptor. The sensor caninclude one or more of a recognition-based substrate, an aptamer-basedsubstrate, an antibody-based substrate, surface plasmon resonance,genetically-modified cells, or genetically-modified cells withreceptor-linked signaling. The genetically-modified cells can includereceptor-linked signaling by fluorogen-activating proteins.

The sensor can be configured to target the device to a site having anelevated level of the inflammatory mediators. The sensor can beconfigured to detect cytokines, T-lymphocytes, B-lymphocytes, orantibodies. The sensor can be configured to detect body temperature,vital signs, edema, oxygen level, or pathogen/toxin level of thesubject. The sensor can be configured to detect one or more ofanaphylatoxin, cytokine, chemokine, leukotriene, prostaglandin,complement, coagulation factor, or proinflammatory cytokine. The sensorcan be configured to detect one or more of TNF-α, IL-1, IL-1β, IL-6,IL-8, IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a,exotoxin, or endotoxin.

The one or more binding agents can be configured to sequester at leastone of the one or more inflammatory mediators from the blood. The atleast one of the one or more inflammatory mediators can be sequesteredby the one or more binding agents prior to treatment with one or more ofa denaturing agent, a degradative agent, binding agent, or an energysource. The one or more binding agents can include, but are not limitedto, one or more of antibodies, receptors, or cognates configured to bindto at least one of the one or more inflammatory mediators. The one ormore binding agent includes, but is not limited to, at least one oflectin, binding protein, catalytic antibody, catalytic aptamer, proteaseconjugate, or photoactivatable agent conjugate. The matrix includes, butis not limited to, one or more of a specific binding ligand or ahydrophobic surface. The matrix can include, but is not limited to, oneor more of beads, cells, vesicles, filters, hydrogel polymers,microparticles, nanoparticles, adsorbent, or synthetic polymers. Thespecific binding ligand or the hydrophobic surface can include, but isnot limited to, one or more of nucleic acid aptamers, peptide aptamers,molecular imprinting polymer, antibodies or fragments thereof, highaffinity mimetics, synthetic binding molecules, or receptor bindingmolecules. The matrix can include, but is not limited to, one or more ofa lectin, binding protein, receptor, antibody, catalytic antibody,catalytic aptamer, protease conjugate, or photoactivatable agentconjugate.

In an aspect, the one or more reactive components can be configured todecrease an activity of one or more of TNF-α, IL-1, IL-1β, IL-6, IL-8,IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin,or endotoxin. The one or more reactive components can modulate anactivity of an intermediate which modulates the activity of one or moreof TNF-α, IL-1, IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, IFN-γ, LIF, MIF,MIP-1, MCP-1, C3-a, C5-a, exotoxin, or endotoxin. The one or morereactive components can increase an activity of an intermediate whichdecreases the activity of one or more of TNF-α, IL-1, IL-1β, IL-6, IL-8,IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin,or endotoxin. The one or more reactive components can decrease anactivity of an intermediate which decreases the activity of one or moreof TNF-α, IL-1, IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, IFN-γ, LIF, MIF,MIP-1, MCP-1, C3-a, C5-a, exotoxin, or endotoxin. In an aspect, the oneor more reactive components can be configured to modulate an activity ofone or more of anaphylatoxins, cytokines, chemokines, leukotrienes,prostaglandins, complement, coagulation factors, or proinflammatorycytokines.

The energy source can include, but is not limited to, acoustic energy orelectronic energy. The energy source can include ultrasound. The energysource can include high-intensity focused ultrasound. The energy sourcecan include, but is not limited to, at least one of microwaveirradiation, gamma irradiation, electron beam irradiation,vibrational/frequency irradiation, or atmospheric pressure glowdischarge. The vibrational/frequency irradiation can include a set ofdiffering energy inputs specifically directed to the one or moreinflammatory mediators, wherein the set of differing energy inputsselectively resonates a plurality of resonant structures in the one ormore inflammatory mediators, and wherein the resonance controllablyalters or reduces the activity of the one or more inflammatory mediatorsin the peripheral blood of the subject. The one or more inflammatorymediators can be modified with a functional group configured to beresponsive to the set of differing energy inputs. The energy source caninclude, but is not limited to, acoustic energy or electronic energy.The one or more denaturing agents can include, but is not limited to, atleast of an acid, base, solvent, cross-linking agent, chaotropic agent,disulfide bond reducer, enzyme, drug, cell, or radical ion. The one ormore degradative agents can include, but is not limited to, at least oneof an enzyme, coenzyme, enzyme complex, catalytic antibody, proteasome,strong acid, strong base, radical, photoactivatable agent, drug, cell,or radical ion. The catalytic antibody can generate a radical ion. Theone or more reactive components can include an activator or an inhibitorof one or more of a myeloid cell, lymphocyte, or precursor thereof. Theone or more reactive components can include an activator or an inhibitorof one or more of a T-lymphocyte, B-lymphocyte, macrophage, monocyte,neutrophil, or NK cell. The one or more reactive components can includea target-specific release compound including one or more cytokines. Thetarget-specific release compound can include DNA aptamer-crosslinkedhydrogel including the one or more cytokines.

In an aspect, the treatment chamber includes a source for producing theone or more reactive components. The source can include at least onereservoir and at least one producer. The at least one producer caninclude at least one encapsulated cell. The at least one encapsulatedcell can produce the one or more reactive components. The at least oneencapsulated cell can be genetically-engineered. The at least oneencapsulated cell can include, but is not limited to, at least one of amammalian cell, bacterial cell, yeast cell, plant cell, insect cell,artificial cell, or enucleated cell. The at least one encapsulated cellcan include, but is not limited to, one or more of a myeloid cell,lymphocyte, or precursor thereof. The at least one encapsulated cell caninclude, but is not limited to, one or more of a T-lymphocyte,B-lymphocyte, macrophage, monocyte, neutrophil, or NK cell. The at leastone producer can include, but is not limited to, a protein, lipidmicelle, liposome, synthetic polymer, or a combination thereof. The atleast one producer can include a catalytic antibody. The catalyticantibody can include a radical ion generator. In an aspect, thetreatment chamber can include one or more reservoirs including one ormore reactive components.

In an aspect, the one or more reactive components can include at leastone modulator of the one or more inflammatory mediators configured toalter the functional structure of the one or more inflammatory mediatorsin the peripheral blood of the subject. The one or more reactivecomponents can include, but is not limited to, at least one of acytokine, cytokine modulator, matrix metalloproteinase (MMP), orextracellular matrix metalloproteinase inducer (EMMPRIN). The at leastone modulator can include an agonist or antagonist of the one or moreinflammatory mediators. The one or more energy source can include anultrasound source to increase cytokine production. The ultrasound sourcecan provide a pulsed radio frequency ultrasound signal.

The one or more inflammatory mediators can include, but is not limitedto, at least one anaphylatoxin, cytokine, chemokine, leukotriene,prostaglandin, complement, coagulation factor, or proinflammatorycytokine. The one or more inflammatory mediators can include, but is notlimited to, one or more of TNF-α, IL-1, L-1β, IL-6, IL-8, IL-10, IL-12,LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin or endotoxin.

In an aspect, the device is intracorporeal. The device can be configuredto be implanted. The device includes, but is not limited to, a stent,bypass implant, nanostructure or microstructure. The device can beconfigured to be implanted relative to an organ or tissue in thesubject. In an aspect, the device is extracorporeal. The extracorporealdevice can include, but is not limited to, a dialysis device,hemoperfusion device, apheresis device, intravenous device, or patchdevice. The inflammatory disease or condition can include, but is notlimited to, one or more of systemic inflammatory response syndrome,sepsis, septic shock, multiple organ dysfunction syndrome, ischemiareperfusion, hyperreactive airway disease, allergic reaction,anaphylaxis, pulmonary failure, adult respiratory distress syndrome(ARDS), allograft rejection, graft versus host disease (GVHD), chronicinflammatory disease, psoriatic arthritis, rheumatoid arthritis,chemical or biological agent exposure, infectious disease, malaria,anthrax, viral hepatitis, viral infection, influenza, smallpox, HIV,myocardial ischemia, or autoimmune disease

A system is disclosed that includes a device including a sensorconfigured to detect one or more inflammatory mediators in peripheralblood of a subject, a controller in communication with the sensor, and ameans for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject. In an aspect, the means for modulating a physiologicaleffect of the one or more inflammatory mediators includes a treatmentchamber configured to receive at least a portion of the peripheral bloodthrough a flow route, the controller configured to control flow ofperipheral blood through the flow route into the treatment chamber, andthe treatment chamber including one or more reactive componentsconfigured to alter a functional structure of the one or moreinflammatory mediators in the peripheral blood. In a further aspect, themeans for modulating a physiological effect of the one or moreinflammatory mediators includes the controller configured to controlinteraction between one or more reactive components and the one or moreinflammatory mediators, the one or more reactive components configuredto alter a functional structure of the one or more inflammatorymediators in the peripheral blood.

A device is disclosed that includes a system including a signal-bearingmedium including one or more instructions for treatment of peripheralblood of a subject through a device including a sensor configured todetect one or more inflammatory mediators in the peripheral blood, oneor more instructions for receiving data including data from a controllerin communication with the sensor, and one or more instructions for ameans for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject. In an aspect, the one or more instructions for the meansfor modulating a physiological effect of the one or more inflammatorymediators includes a treatment chamber configured to receive at least aportion of the peripheral blood through a flow route, the controllerconfigured to control flow of peripheral blood through the flow routeinto the treatment chamber, and the treatment chamber including one ormore reactive components configured to alter a functional structure ofthe one or more inflammatory mediators in the peripheral blood. In afurther aspect, the one or more instructions the means for modulating aphysiological effect of the one or more inflammatory mediators includesthe controller configured to control interaction between one or morereactive components and the one or more inflammatory mediators, the oneor more reactive components configured to alter a functional structureof the one or more inflammatory mediators in the peripheral blood.

A system is disclosed that includes at least one computer programincluded on a computer-readable medium for use with at least onecomputer system wherein the computer program includes a plurality ofinstructions including one or more instructions for treatment ofperipheral blood of a subject through a device including a sensorconfigured to detect one or more inflammatory mediators in theperipheral blood, one or more instructions for receiving data includingdata from a controller in communication with the sensor, and one or moreinstructions for a means for modulating a physiological effect of theone or more inflammatory mediators responsive to the controller, thecontroller configured to adjust the modulating means to achieve a targetvalue of the detected one or more inflammatory mediators in theperipheral blood of the subject. In an aspect, the one or moreinstructions for the means for modulating a physiological effect of theone or more inflammatory mediators includes a treatment chamberconfigured to receive at least a portion of the peripheral blood througha flow route, the controller configured to control flow of peripheralblood through the flow route into the treatment chamber, and thetreatment chamber including one or more reactive components configuredto alter a functional structure of the one or more inflammatorymediators in the peripheral blood. In a further aspect, the one or moreinstructions the means for modulating a physiological effect of the oneor more inflammatory mediators includes the controller configured tocontrol interaction between one or more reactive components and the oneor more inflammatory mediators, the one or more reactive componentsconfigured to alter a functional structure of the one or moreinflammatory mediators in the peripheral blood.

The foregoing summary is illustrative only and is not intended to be inany way limiting. In addition to the illustrative aspects, embodiments,and features described above, further aspects, embodiments, and featureswill become apparent by reference to the drawings and the followingdetailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B depict a diagrammatic view of an aspect of an exemplaryembodiment of an intracorporeal device.

FIGS. 2A and 2B depict a diagrammatic view of an aspect of an exemplaryembodiment of an intracorporeal device.

FIGS. 3A and 3B depict a diagrammatic view of an aspect of an exemplaryembodiment of an extracorporeal device.

FIG. 4 depict a diagrammatic view of an aspect of an exemplaryembodiment of an intracorporeal device.

FIG. 5 depict a diagrammatic view of an aspect of an exemplaryembodiment of a device.

FIG. 6 depicts a logic flowchart of a method for a method for treatingan inflammatory disease or condition in a subject.

FIG. 7 depicts a logic flowchart of a method for treating aninflammatory disease or condition in a subject

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description and drawings are not meant to be limiting. Otherembodiments may be utilized, and other changes may be made, withoutdeparting from the spirit or scope of the subject matter presented here.

The present application uses formal outline headings for clarity ofpresentation. However, it is to be understood that the outline headingsare for presentation purposes, and that different types of subjectmatter may be discussed throughout the application (e.g., method(s) maybe described under composition heading(s) and/or kit headings; and/ordescriptions of single topics may span two or more topic headings).Hence, the use of the formal outline headings is not intended to be inany way limiting.

Devices, systems, and methods are described herein for controlling ormodulating an inflammatory response in a subject. Devices, systems, andmethods are disclosed that alter the functional structure of one or moreinflammatory mediators in the peripheral blood of the subject. Devices,systems, and methods are disclosed that alter the biological activity ofone or more inflammatory mediators in the peripheral blood of thesubject The device or system described herein is useful in a method fortreating an inflammatory disease or condition in the subject. Diseasesor conditions related to either acute or chronic inflammatory responseinclude, but are not limited to, systemic inflammatory responsesyndrome, sepsis, septic shock, multiple organ dysfunction syndrome,ischemia reperfusion, hyperreactive airway disease, (e.g., asthma,chronic obstructive pulmonary disease, rhinitis, sinusitis), allergicreaction, anaphylaxis, pulmonary failure, adult respiratory distresssyndrome (ARDS), allograft rejection, graft versus host disease (GVHD),chronic inflammatory disease, psoriatic arthritis, rheumatoid arthritis,chemical or biological agent exposure, infectious disease, malaria,anthrax, viral hepatitis, viral infection, influenza, smallpox, HIV,myocardial ischemia, or autoimmune disease.

A device is disclosed that includes a sensor configured to detect one ormore inflammatory mediators in peripheral blood of a subject, acontroller in communication with and responsive to the sensor, and ameans for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject. The means for modulating a physiological effect of theone or more inflammatory mediators can include a treatment chamberconfigured to receive at least a portion of the peripheral blood througha flow route, the controller configured to control flow of peripheralblood through the flow route into the treatment chamber, and thetreatment chamber including one or more reactive components configuredto alter a functional structure of the one or more inflammatorymediators in the peripheral blood. The controller can be configured tocontrol access to the treatment chamber by the peripheral blood,including access to one or more reactive components in the treatmentchamber. The one or more reactive components can include, but are notlimited to, one or more of a denaturing agent, a degradative agent,binding agent, or an energy source. The one or more binding agents on amatrix adapted to the treatment chamber can be configured to sequesterat least one of the one or more inflammatory mediators from the blood.The at least one of the one or more inflammatory mediators can besequestered by the binding agent prior to treatment with one or more ofa denaturing agent, a degradative agent, binding agent, or an energysource.

In a further aspect, the means for modulating a physiological effect ofthe one or more inflammatory mediators can include the controllerconfigured to control interaction between one or more reactivecomponents and the one or more inflammatory mediators, the one or morereactive components configured to alter a functional structure of theone or more inflammatory mediators in the peripheral blood. Thecontroller can be in communication with a reservoir configured torelease the one or more reactive components into the peripheral blood ofthe subject.

The modulating means can be configured to altering a functionalstructure of the one or more inflammatory mediators which refers todecreasing an activity of the one or more of inflammatory mediators, forexample, by denaturation, degradation, or inactivation by one or more ofan energy source, a biological agent, or a chemical agent. The one ormore reactive components can include, but are not limited to, one ormore of a denaturing agent, a degradative agent, a binding agent, amodulator, or an energy source. The one or more binding agents caninclude one or more of antibodies, receptors, or cognates configured tobind to at least one of the one or more inflammatory mediators.

In an aspect, the target value can include a desired concentration ofthe one or more inflammatory mediators in the peripheral blood. Thetarget value can include a desired range of concentrations of the one ormore inflammatory mediators in the peripheral blood. The target valuecan include a desired ratio of concentrations of two or moreinflammatory mediators in the peripheral blood. The sensor and thecontroller can be configured to control levels of the one or moreinflammatory mediators to substantially attain the target value. Thesensor and the controller can be configured to control levels of the oneor more inflammatory mediators to limit a deviation from the targetvalue. The deviation can be determined by a weighted least squares fit.

A method for treating an inflammatory disease or condition in a subjectincludes providing a device configured to communicate with at least aportion of a peripheral blood of the subject, the device including asensor configured to detect one or more inflammatory mediators inperipheral blood of a subject, a controller in communication with andresponsive to the sensor, a means for modulating a physiological effectof the one or more inflammatory mediators responsive to the controller,the controller configured to adjust the modulating means to achieve atarget value of the detected one or more inflammatory mediators in theperipheral blood of the subject.

A system is disclosed that includes a device including a sensorconfigured to detect one or more inflammatory mediators in peripheralblood of a subject, a controller in communication with and responsive tothe sensor, and a means for modulating a physiological effect of the oneor more inflammatory mediators responsive to the controller, thecontroller configured to adjust the modulating means to achieve a targetvalue of the detected one or more inflammatory mediators in theperipheral blood of the subject.

Inflammation, Inflammatory Mediators and Inflammatory Disease orCondition

Inflammation is a complex biological response to insults that arisefrom, for example, chemical, traumatic, or infectious stimuli. It is aprotective attempt by an organism to isolate and eradicate the injuriousstimuli as well as to initiate the process of tissue repair. The eventsin the inflammatory response are initiated by a complex series ofinteractions involving inflammatory mediators, including those releasedby immune cells and other cells of the body. Histamines and eicosanoidssuch as prostaglandins and leukotrienes act on blood vessels at the siteof infection to localize blood flow, concentrate plasma proteins, andincrease capillary permeability. Chemotactic factors, including certaineicosanoids, complement, and especially cytokines known as chemokines,attract particular leukocytes to the site of infection. Otherinflammatory mediators, including some released by the summonedleukocytes, function locally and systemically to promote theinflammatory response. Platelet activating factors and related mediatorsfunction in clotting, which aids in localization and can trap pathogens,Certain cytokines, e.g., tumor necrosis factor (TNF), and interleukinsinduce further trafficking and extravasation of immune cells,hematopoiesis, fever, and production of acute phase proteins. Oncesignaled, some cells and/or their products directly affect the offendingpathogens, for example by inducing phagocytosis of bacteria or, as withinterferon, providing antiviral effects by shutting down proteinsynthesis in the host cells. Oxygen radicals, cytotoxic factors andgrowth factors may also be released to fight pathogen infection and/orto facilitate tissue healing. This cascade of biochemical eventspropagates and matures the inflammatory response, involving the localvascular system, the immune system, and various cells within the injuredtissue. Under normal circumstances, through a complex process ofmediator-regulated pro-inflammatory and anti-inflammatory signals, theinflammatory response eventually resolves itself and subsides. Forexample, the transient and localized swelling associated with a cut isan example of an acute inflammatory response. However, in certain casesresolution may not occur as expected. Prolonged inflammation, known aschronic inflammation, may lead to a progressive shift in the type ofcells present at the site of inflammation and is characterized bysimultaneous destruction and healing of the tissue from the inflammatoryprocess, as directed by certain mediators. Rheumatoid arthritis is anexample of a disease associated with persistent and chronicinflammation.

In some aspects, the inflammatory response becomes uncontrolled as isthe case with systemic immune response syndrome (SIRS). SIRS is aninflammatory state of the entire body and may be initiated by ischemia,inflammation, trauma, infection, or a combination thereof. A triggeringevent, such as trauma, may induce localized release of inflammatorymediators with the goal of initiating an inflammatory response topromote wound repair. Small amounts of local inflammatory mediators arereleased into the circulation to improve the local response. This maylead to growth factor stimulation and recruitment of macrophages andplatelets. Such an acute phase response may be typically well-controlledby a natural decrease in endogenous pro-inflammatory mediators and bythe release of endogenous antagonists with the goal of restoringhomeostasis. However, if homeostasis is not restored, a significantsystemic reaction may occur leading to what is known as “cytokine storm”or hypercytokinemia. A consequence of this is the activation of numeroushumoral cascades, the activation of the reticular endothelial system andthe subsequent loss of circulatory integrity, potentially leading toend-organ dysfunction and death. In this aspect, the excessive releaseof inflammatory mediators may lead to destruction of cells and tissuerather than protection. Hypercytokinemia has the potential to dosignificant damage to body tissues and organs.

Both pro-inflammatory and anti-inflammatory mediators are released intothe peripheral system during the hypercytokinemia associated with SIRS.In the case of sepsis or infection-induced SIRS, bacterial ligands suchas lipopolysaccharide (LPS) activate toll-like receptors (TLRs) andNF-κB, leading to increased expression and release of thepro-inflammatory mediators, e.g., tumor necrosis factor α (TNF-α) andinterleukins IL-1, IL-6, IL-8, and IL-12. See, e.g., Sriskandan &Altmann, J. Pathol. 214:211-233, 2008, which is incorporated herein byreference. In contrast, anti-inflammatory mediators such as interleukinsIL-4, IL-10 and IL-13 as well as transforming growth factor β (TGF-B-β)suppress gene expression and the synthesis of IL-1, TNF and otherpro-inflammatory cytokines. See, e.g., Dinarello Chest 112:321-329,1997, which is incorporated herein by reference. The inability of theimmune system to return to homeostasis by balancing the pro-inflammatoryand anti-inflammatory mediators may contribute to progression of SIRS tomultiple organ failure and potentially death.

Inhibition of TNF-α or IL-1 activity with selective antagonists has notbeen a successful approach for treating patients with sepsis/SIRS. See,e.g., Remick Curr. Pharm. Des. 9:75-82, 2003). Non-selective removal ofinflammatory mediators using hemodialysis, hemofiltration,hemoadsorption, or plasma filtration has had limited success. See, e.g.,Venkataraman, et al., Critical Care 7:139-145, 2003; Kushi, et al.,Critical Care 9:R659-661, 2005; Nakakda, et al., Transfus. Apher. Sci.35:253-264, 2006; Nakada, et al., Mol. Med. 14:257-263, 2008, each ofwhich is incorporated herein by reference. In the non-selectiveapproach, both pro-inflammatory and anti-inflammatory mediators areremoved from the blood.

Another approach to treating the severe inflammatory response associatedwith an inflammatory condition can involve controllably removing,modulating or inactivating specific inflammatory mediators at differenttimes over the course of the inflammatory response. The controllableremoval, modulation, or inactivation of specific inflammatory mediatorsmay be based on real-time monitoring of a subject's blood. The removal,modulation, or inactivation of at least one inflammatory mediatorincludes, for example, altering the functional structure of the at leastone inflammatory mediator accomplished by blocking a binding site oractive site of the at least one inflammatory mediator, by modulating theexpression of the at least one inflammatory mediator, by modulating thephysiological effect, or by agonizing the activity or expression of ananti-inflammatory mediator.

A device is described herein for altering the functional structure ofone or more inflammatory mediators in the peripheral blood of a subject.The modulating means can be configured to altering a functionalstructure of the one or more inflammatory mediators which refers todecreasing an activity of the one or more of inflammatory mediators, forexample, by denaturation, degradation, or inactivation by one or more ofan energy source, a biological agent, or a chemical agent. The devicecan be used in a method for treating an inflammatory disease orcondition in the subject. The device can be extracorporeal orintracorporeal, or a combination thereof. The device can include one ormore sensors configured to detect one or more inflammatory mediators inthe peripheral blood of a subject and configured to control levels ofthe one or more inflammatory mediators to a target value. The one ormore sensors can include, for example, a biosensor, a chemical sensor, aphysical sensor, an optical sensor, or a combination thereof. The devicecan further include one or more treatment chambers configured to receivethe peripheral blood of a subject through a flow route. The one or moretreatment chambers can include one or more specific binding agents forbinding one or more specific inflammatory mediators. The one or morespecific binding agents can be attached to one or more substrates in theone or more treatment chambers. The one or more substrates can be one ormore surfaces of the one or more treatment chambers. The one or moresubstrates can include one or more matrix components retained in the oneor more treatment chambers. The one or more treatment chambers caninclude one or more reactive components configured to alter thefunctional structure of one or more inflammatory mediators found in theperipheral blood of a subject that flow through the flow route. The oneor more reactive components include, but are not limited to, adenaturing agent, a degradative agent, an energy source, or acombination thereof. The device further includes a controller thatreceives data from the one or more sensors and in response to the inputdata controls flow into and out of the one or more treatment chambersand/or controls release and/or activation of one or more reactivecomponents.

In a further aspect, a method is provided for treating an inflammatorydisease or condition in a subject by providing a device configured tocommunicate with the peripheral blood of the subject. One or moresensors are configured to detect one or more inflammatory mediators inthe peripheral blood of the subject. In response to sensing one or moreinflammatory mediators, a flow route is provided from the peripheralblood to one or more treatment chambers. The one or more treatmentchambers are configured to alter the functional structure of the one ormore inflammatory mediators in the peripheral blood and to modulate,reduce, alleviate, or eliminate the inflammatory disease or condition inthe subject.

Inflammatory diseases or conditions wherein the inflammation associatedwith the disease or condition may be modulated, alleviated, treated,prevented, reduced or eliminated by altering the functional structure ofone or more inflammatory mediators in the peripheral blood include, butare not limited to, systemic inflammatory response syndrome, sepsis,septic shock, multiple organ dysfunction syndrome, ischemia reperfusion,hyperreactive airway disease, (e.g., asthma, chronic obstructivepulmonary disease, rhinitis, sinusitis), allergic reaction, anaphylaxis,pulmonary failure, adult respiratory distress syndrome (ARDS), allograftrejection, graft versus host disease (GVHD), chronic inflammatorydisease, psoriatic arthritis, rheumatoid arthritis, chemical orbiological agent exposure (due to warfare, accident, or occupation),infectious disease, malaria, anthrax, viral infection, viral hepatitis,influenza, smallpox, acquired immunodeficiency syndrome (AIDS)associated with HIV infection, myocardial ischemia, or autoimmunedisease.

Inflammatory mediators as used herein include pro-inflammatory mediatorsor anti-inflammatory mediators, or both. The types of inflammatorymediators that may be modulated, agonized, antagonized, reduced, oreliminated by altering, for example, their biological activity orexpression or the functional structure include, but are not limited to,cytokines such as interferons (IFN) IFN-α, IFN-β, and IFN-γ;interleukins (IL) IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9,IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19,IL-20, IL-21, IL-22, IL-23, IL-24, IL-27, IL-28, IL-29, IL-30, IL-31,and IL-32; tumor necrosis factor (TNF) TNF-α and TNF-β; granulocytecolony stimulating factor (G-CSF); granulocyte-macrophage colonystimulating factor (GM-CSF); macrophage colony-stimulating factor(M-CSF); erythropoietin (EPO); and thrombopoietin (TPO). The one or moreinflammatory mediators may be any of a number of chemotactic cytokines(chemokines) including but not limited to CC chemokines CCL1 throughCCL28 exemplified by RANTES (CCL5), MCP-1 (CCL2), LARC(CCL20), MIP-1 α(CCL3), and MDC (CCL22); CXC chemokines CXCL1 through CXCL17 exemplifiedby LIX (CXCL5), GCP-2 (CXCL6) and BCA-1 (CXCL13); C chemokines XCL1 andXCL2; CX3C chemokine C3CL1 (fractalkine); and chemokine like moleculesexemplified by MIF. Other inflammatory mediators include but are notlimited to anaphylatoxin fragments C3a, C4a, and C5a from the complementpathway; leukotrienes LTA4, LTB4, LTC4, LTD4, LTE4, and LTF4;prostaglandins; growth factors EGF, FGF-9, FGF-basic, growth hormone,stem cell factor (SCF), TGF-13 and VEGF; soluble tumor necrosis factorreceptor (sTNFR I and II); soluble interleukin receptors sIL-1r andsIL-2r; C-reactive protein; CD11b; histamine; serotonin; apolipoproteinA1; β2-microglobulin; bradykinin; D-dimer; endothelin-1; eotaxin; factorVII; fibrinogen; GST; haptoglobin; IgA; insulin; IP-10; leptin; LIF;lymphotactin; myoglobin; OSM; SGOT; TIMP-1; tissue factor; VCAM-1; VWF;thromboxane; platelet activating factor (PAF); immunoglobulins; andpathogen-derived products including endotoxins such aslipopolysaccharide (LPS); and various exotoxins such as superantigens,e.g., from Staphylococcus aureus and Streptococcus pyogenes.

Anti-inflammatory mediators include, but are not limited to, IL-4,IL-10, IL-13, transforming growth factor-β (TGF-β), endogenous IL-1receptor antagonists, as well as endogenous soluble TNF receptors,cyclopentenone prostaglandin D₂ (PGD₂), PGE₂, annexin-1, galectin-1,interferon-α, interferon-β, and lipoxins. IL-4 and IL-13 may beconsidered anti-inflammatory mediators because they block production ofIL-6, IL-1 and TNF-α but they also upregulate the cell surface receptorfor IgE complexes, CD23, and stimulate B cells to stimulate IgEproduction. IL-10 also blocks IL-1 and TNF.

Pro-inflammatory mediators may include, but are not limited to, TNF-α,IL-113, IL-6, and IL-8, prostaglandins (e.g., PGE2), leukotrienes (e.g.,LTB4), CXC chemokines (e.g., macrophage inflammatory protein-2), andplatelet activating factor Interleukin-6 (IL-6) is an interleukin thatmay act as both a pro-inflammatory and anti-inflammatory cytokine. IL-6is secreted by T cells and macrophages to stimulate immune response totrauma, especially burns or other tissue damage leading to inflammation.The role of IL-6 as an anti-inflammatory cytokine is mediated throughits inhibitory effects on TNF-α and IL-1, and activation of IL-1ra andIL-10.

With reference to the figures, and with reference now to FIGS. 1, 2, 3and 4, depicted is an aspect of a device, system, or method that mayserve as an illustrative environment of and/or for subject mattertechnologies, for example, a device including a sensor configured todetect one or more inflammatory mediators in peripheral blood of asubject and configured to control levels of the one or more inflammatorymediators to a target value, and a treatment chamber responsive tooutput from the sensor and configured to receive the peripheral bloodthrough a flow route, the treatment chamber including one or morereactive components configured to alter a functional structure of theone or more inflammatory mediators in the peripheral blood of thesubject. The device may be intracorporeal or extracorporeal of thesubject. The specific methods described herein are intended as merelyillustrative of their more general counterparts.

Continuing to refer to FIGS. 1A and 1B, depicted is a partialdiagrammatic view of an illustrative embodiment of a device 100including a sensor 110 configured to detect one or more inflammatorymediators 150 in peripheral blood 160 of a subject and configured tocontrol levels of the one or more inflammatory mediators to a targetvalue, and a treatment chamber 120 including one or more reactivecomponents 130 configured to alter a functional structure of the one ormore inflammatory mediators 150 in the peripheral blood of the subject.In FIG. 1A, the device can further include a controller 140 incommunication with and responsive to the sensor 110 and configured tocontrol access to the treatment chamber 120 by the peripheral blood 160.The reactive component can further include an affinity binding component170 to bind the one or more inflammatory mediators 150 and configured toalter a functional structure of the one or more inflammatory mediators.The one or more reactive components 130 include, but are not limited to,a denaturing agent, a degradative agent, or an energy source, which candestroy, degrade, denature the one or more inflammatory mediators 150 ordestroy an activity of the one or more inflammatory mediators. Thedevice can be configured to reduce the concentration of inflammatorymediators at one or more sites of inflammation 180 in the subject, orthroughout the peripheral blood.

In FIG. 1B, the device can further include a controller 140 configuredto be in communication with and responsive to the sensor 110 andconfigured to control access to the treatment chamber 120 by theperipheral blood 160. The reactive component can further include adiffusible component 170 to bind the one or more inflammatory mediators150 and configured to alter a functional structure of the one or moreinflammatory mediators. The diffusible component 170 can bind to the oneor more inflammatory mediators 150 within the treatment chamber oroutside in the peripheral blood of the subject. The one or more reactivecomponents 130 or the diffusible component 170 include, but are notlimited to, a denaturing agent, a degradative agent, or an energysource, which can destroy, degrade, denature the one or moreinflammatory mediators 150 or destroy an activity of the one or moreinflammatory mediators.

Continuing to refer to FIGS. 2A and 2B, depicted is a partialdiagrammatic view of an illustrative embodiment of a device 200including a sensor 210 configured to detect one or more inflammatorymediators 250 in peripheral blood 260 of a subject and configured tocontrol levels of the one or more inflammatory mediators to a targetvalue, and a treatment chamber 220 including one or more reactivecomponents 230 configured to alter a functional structure of the one ormore inflammatory mediators 250 in the peripheral blood of the subject.In FIG. 2A, the device can further include a controller 240 incommunication with and responsive to the sensor 210 and configured tocontrol access to the treatment chamber 220 by the peripheral blood 260.The one or more reactive component can further include a diffusibleagent 270 to bind a precursor or inflammatory modulator 290 and/orinhibit synthesis of the one or more inflammatory mediators 250. The oneor more reactive components 230 include, but are not limited to, adenaturing agent, a degradative agent, or an energy source, which candestroy, degrade, denature the one or more inflammatory mediators 250 ordestroy an activity of the one or more inflammatory mediators. Thedevice includes the sensor configured to detect a site having anelevated level of the inflammatory mediators. The device can be targetedto a site having elevated levels in order to reduce the concentration ofinflammatory mediators at one or more sites of inflammation 280 in thesubject.

In FIG. 2B, the device can further include a controller 240 incommunication with and responsive to the sensor 210 and configured tocontrol access to the treatment chamber 220 by the peripheral blood 260.The one or more reactive component can further include an encapsulatedcell that produces a diffusible agent 270 configured to bind a precursor290 and/or inhibit synthesis of the one or more inflammatory mediators250 and configured to alter a functional structure of the one or moreinflammatory mediators. In an aspect, the reactive component can includean encapsulated cell that produces a diffusible agent 270 configured tobind to the one or more inflammatory mediators 250 and configured toalter a functional structure of the one or more inflammatory mediators.The one or more reactive components 230 include, but are not limited to,a denaturing agent, a degradative agent, or an energy source, which candestroy, degrade, denature the one or more inflammatory mediators 250 ordestroy an activity of the one or more inflammatory mediators. Thedevice is configured to reduce the concentration of inflammatorymediators at one or more sites of inflammation 280 in the subject.

Continuing to refer to FIGS. 3A and 3B, depicted is a partialdiagrammatic view of an illustrative embodiment of a device 300including a sensor 310 configured to detect one or more inflammatorymediators 350 in peripheral blood 360 of a subject and configured tocontrol levels of the one or more inflammatory mediators to a targetvalue, and a treatment chamber 320 including one or more reactivecomponents 330 configured to alter a functional structure of the one ormore inflammatory mediators 350 in the peripheral blood of the subject.The device is outside the peripheral blood vessel of the subject and canbe configured as an extracorporeal device.

In FIG. 3A, the device 300 can further include a controller 340 incommunication with and responsive to the sensor 310 and configured tocontrol access to the treatment chamber 320 by the peripheral blood 360.The reactive component 330 can further include a diffusible component370 to bind the one or more inflammatory mediators 350 and configured toalter a functional structure of the one or more inflammatory mediators.The one or more reactive components 330 or the diffusible component 370include, but are not limited to, a denaturing agent, a degradativeagent, or an energy source, which can destroy, degrade, denature the oneor more inflammatory mediators 350 or destroy an activity of the one ormore inflammatory mediators. The device is configured to reduce theconcentration of inflammatory mediators at one or more sites ofinflammation 380 in the subject.

In FIG. 3B, the device 300 can further include a controller 340 incommunication with and responsive to the sensor 310 and configured tocontrol access to the treatment chamber 320 by the peripheral blood 360.The reactive component 330 can further include an affinity bindingcomponent 370 to bind the one or more inflammatory mediators 350 andconfigured to alter a functional structure of the one or moreinflammatory mediators. The one or more reactive components 330 include,but are not limited to, a denaturing agent, a degradative agent, or anenergy source, which can destroy, degrade, denature the one or moreinflammatory mediators 350 or destroy an activity of the one or moreinflammatory mediators. The device is configured to reduce theconcentration of inflammatory mediators at one or more sites ofinflammation 380 in the subject.

Continuing to refer to FIG. 4, depicted is a partial diagrammatic viewof an illustrative embodiment of a device 400 including a sensor 410configured to detect one or more inflammatory mediators 450 inperipheral blood 460 of a subject and configured to control levels ofthe one or more inflammatory mediators to a target value, and areservoir 470 including one or more reactive components 430 configuredto alter a functional structure of the one or more inflammatorymediators 450 in the peripheral blood of the subject. The device canfurther include a controller 440 in communication with and responsive tothe sensor 410. The reactive component 430 can further include adiffusible component to bind the one or more inflammatory mediators 450and configured to alter a functional structure of the one or moreinflammatory mediators. The one or more reactive components 430 include,but are not limited to, a denaturing agent, a degradative agent, or anenergy source, which can destroy, degrade, denature the one or moreinflammatory mediators 450 or destroy an activity of the one or moreinflammatory mediators. The device can be configured to reduce theconcentration of inflammatory mediators at one or more sites ofinflammation 480 in the subject, or throughout the peripheral blood.

Referring to FIG. 5, depicted is a partial diagrammatic view of anillustrative embodiment of a device including a sensor configured todetect one or more inflammatory mediators in peripheral blood of asubject and configured to control levels of the one or more inflammatorymediators to a target value, and a treatment chamber responsive tooutput from the sensor and configured to receive the peripheral bloodthrough a flow route, the treatment chamber including one or morereactive components configured to alter a functional structure of theone or more inflammatory mediators in the peripheral blood of thesubject. In an aspect, the target value includes a desired concentrationof the one or more inflammatory mediators in the peripheral blood, orthe target value includes a desired range of concentrations of the oneor more inflammatory mediators in the peripheral blood. In a furtheraspect, the target value includes a desired ratio of concentrations oftwo or more inflammatory mediators in the peripheral blood. Or it candetermine relative levels of the inflammatory mediators. The desiredratio of concentrations can be determined by any method or means,including for example, by a least squares fit of the concentrations ofthe two or more inflammatory mediators. For example, the desired ratioof concentrations can be determined by a least squares fit of theconcentrations of the two or more inflammatory mediators atconcentrations x₁, x₂, x₃, and x₄ for a first inflammatory mediator, X,and at concentrations y₁, y₂, y₃, and y₄ for a second inflammatorymediator, Y. The least squares can fit to a line or to a two or threedimensional space indicating the preferred ratio of the two or moreinflammatory mediators.

Referring to FIG. 6, depicted is a logic flowchart of a method fortreating an inflammatory disease or condition in a subject. The method601 includes 602 providing a device configured to communicate with atleast a portion of a peripheral blood of the subject, the deviceincluding a sensor configured to detect one or more inflammatorymediators in peripheral blood of a subject; a controller incommunication with the sensor, a means for modulating a physiologicaleffect of the one or more inflammatory mediators responsive to thecontroller, the controller configured to adjust the modulating means toachieve a target value of the detected one or more inflammatorymediators in the peripheral blood of the subject. The means formodulating a physiological effect of the one or more inflammatorymediators includes 604 a treatment chamber configured to receive atleast a portion of the peripheral blood through a flow route, thecontroller configured to control flow of peripheral blood through theflow route into the treatment chamber, and the treatment chamberincluding one or more reactive components configured to alter afunctional structure of the one or more inflammatory mediators in theperipheral blood. The means for modulating a physiological effect of theone or more inflammatory mediators includes 605 the controllerconfigured to control interaction between one or more reactivecomponents and the one or more inflammatory mediators, the one or morereactive components configured to alter a functional structure of theone or more inflammatory mediators in the peripheral blood.

Referring to FIG. 7, depicted is a logic flowchart of a method fortreating an inflammatory disease or condition in a subject. The method701 includes providing an extracorporeal device 702 including atreatment chamber configured to receive peripheral blood of the subjectthrough a flow route, the treatment chamber including one or morereactive biological or chemical compounds that alter the functionalstructure of one or more inflammatory mediators in the peripheral blood.The method further includes 704 providing a sensor configured to detectthe one or more inflammatory mediators in the peripheral blood. Themethod further includes 705 providing a controller in communication withthe sensor and configured to adjust the one or more reactive biologicalor chemical compounds to achieve a target value of the detected one ormore inflammatory mediators in the peripheral blood of the subject.

Controlling Levels of One or More Inflammatory Mediators to a TargetValue

A device is described herein that includes a sensor configured to detectone or more inflammatory mediators in peripheral blood of a subject andconfigured to control levels of the one or more inflammatory mediatorsto a target value. As described above, the target value can be a desiredconcentration of one or more inflammatory mediators in the peripheralblood, or the target value can be a desired range of concentrations ofone or more inflammatory mediators in the peripheral blood.Alternatively, the target value can be a desired ratio of concentrationsof two or more inflammatory mediators in the peripheral blood. Thedesired ratio can be determined by a least squares fit of theconcentrations of the two or more inflammatory mediators. The targetvalue of an inflammatory mediator can be a desired concentration and/orconcentration range and/or ratio of concentrations that is a specificvalue or range of values such as, for example, a value or range ofvalues observed in a normal subject. Alternatively, the target value ofan inflammatory mediator can be a desired concentration and/orconcentration range and/or ratio of concentrations that is at least 20%,at least 40%, at least 60%, at least 80%, or at least 100% below orabove the current level of the inflammatory mediator in the peripheralblood of a subject.

The target value of one or more inflammatory mediators can be a desiredconcentration and/or concentration range that is below that observed inthe peripheral blood of a subject experiencing an inflammatory response,condition, disorder, or disease. A number of inflammatory mediators areelevated in the peripheral blood of subjects diagnosed with systemicimmune response syndrome (SIRS) and sepsis. See, e.g., Ueda, et al., Am.J. Respir. Crit. Care Med. 160:132-136, 1999; Kurt, et al., MediatorsInflamm. 2007:31397, 2007; Kellum, et al., Arch. Intern. Med.167:1655-1663, 2007; Wang, et al., Crit. Care 12:R106, 2008; which areincorporated herein by reference. One study compared the levels ofTNF-α, IL-6, and IL-8 in normal subjects and subjects diagnosed withseptic shock. In this study, the normal ranges of TNF-α, IL-6, and IL-8were reported as less than 5 pg/ml, less than 10 pg/ml, and less than 10pg/ml, respectively. In individuals with septic shock, the serum levelsof TNF-α, IL-6, and IL-8 were significantly elevated to mean values of138+/−22 pg/ml, 27,255+/−7,895 pg/ml, and 2,491+/−673 pg/ml,respectively. Ueda, et al., Am. J. Respir. Crit. Care Med. 160:132-136,1999; which is incorporated herein by reference. As another example, astudy described elevated serum levels of IL-1β in neonates diagnosedwith sepsis versus IL-1β serum levels in normal neonates (41.2+/−13.6pg/ml versus 10.4+/−2.7 pg/ml). In this same study the levels of TNF-α(21.0+/−9.4 pg/ml versus 4.6+/−1.5 pg/ml), IL-6 (196.9+/−74.1 pg/mlversus 8.2+/−5.8 pg/ml), and IL-8 (481.3+/−186.6 pg/ml versus55.5+/−26.5 pg/ml) were also elevated in septic versus non-septicneonates. IL-10 is also present in elevated levels in the peripheralblood of subjects diagnosed with sepsis versus normal controls (39-100pg/ml versus less than 10 pg/ml. Kurt, et al., Mediators Inflamm.2007:31397, 2007; Kellum, et al., Arch. Intern. Med. 167: 1655-1663,2007; which are incorporated herein by reference.

The relative levels of one or more inflammatory mediators in theperipheral blood of a subject may be correlated with prognosis andsurvival. For example, sepsis non-survivors have proportionally higherlevels of inflammatory mediators relative to sepsis survivors and normalcontrols. In one study, high levels of both IL-10 and IL-6 areassociated with increased mortality with a hazard ratio of 20.52. Inthis study, high initial serum levels of IL-10 (mean of 45 pg/ml) andIL-6 (mean of 735 pg/ml) upon admission to the emergency department wereassociated with development of severe sepsis and increased risk of deathas compared with low initial levels of IL-10 (mean of 7.4 pg/ml) andIL-6 (mean of 15 pg/ml). Similarly, a second study demonstrated a directcorrelation between sepsis symptom scores and the serum level of IL-6and suggested that persistent elevation in IL-6 levels is predictive ofpoor outcome. These data suggest that modulating the levels of one ormore inflammatory mediators to a desired target value in the peripheralblood may alter the course of the disease. See, e.g., Kellum, et al.,Arch. Intern. Med. 167:1655-1663, 2007; Presterl, et al., Am. J. Respir.Crit. Care Med. 156:825-832, 1997; which are incorporated herein byreference.

In some instances, the target value of one or more inflammatorymediators may be a desired concentration and/or concentration range thatis above that observed in the peripheral blood of a subject experiencingan inflammatory response, condition, disorder, or disease. For example,subjects with severe sepsis have decreased serum levels of gelsolin, aprotein involved in severing and scavenging circulating filamentousactin. Studies have shown that actin may enhance major components ofproinflammatory cytokine production, impair microcirculation andcompromise multiple organs. Wang, et al., Crit. Care 12:R106, 2008,which is incorporated herein by reference. The normal level of gelsolin(126.8+/−32 mg/l) is depressed in non-septic critically ill subjects(52.3+/−20.3 mg/l) and further depressed in subjects diagnosed withsevere sepsis (20.6+/−11.7 mg/l). In a further example, elevated levelsof IL-10 in patients with acute coronary syndromes and elevatedC-reactive protein are at reduced risk of death relative to similarpatients with lower levels of IL-10. See, e.g., Heeschen, et al., Circ.107: 2109-2114, 2003, which is incorporated herein by reference. Inthese instances, the desired target value may be higher than the levelsensed in the peripheral blood of a subject experiencing an inflammatoryresponse, condition, disorder, or disease.

Arthritis is a chronic inflammatory disease in which changes in theserum levels of various inflammatory mediators have been observed. Forexample, one study compared the mean levels of various inflammatorymediators in normal subjects versus subjects with rheumatoid arthritisincluding IL-6 (4.0 pg/ml versus 15.8 pg/ml), TNFα (3.2 pg/ml versus 10pg/ml), IL-1β (57.3 pg/ml versus 129.8 pg/ml), IL-8 (2.6 pg/ml versus9.3 pg/ml), IL-10 (4.6 pg/ml versus 15.5 pg/ml), and IL-12 (6.2 pg/mlversus 20.2 pg/ml). Psoriatic arthritis is also characterized byincreased levels of circulating inflammatory mediators. Another studycompared the levels of a number of cytokines, chemokines and growthfactors in the serum of subjects diagnosed with psoriatic arthritis.Most notable were statistically significant increases in the serumlevels of various inflammatory mediators in subjects diagnosed withpsoriatic arthritis versus normal controls including IFN-α (38 pg/mlversus 8 pg/ml), IL-10 (14 pg/ml versus 11 pg/ml), IL-13 (11 pg/mlversus 8.5 pg/ml), EGF (80 pg/ml versus 32 pg/ml), VEGF (49 pg/ml versus13 pg/ml), FGF (43 pg/ml versus 16 pg/ml) CCL3 (200 pg/ml versus 45pg/ml) and CCL4 (140 pg/ml versus 50 pg/ml). The range of normal levelsof each inflammatory mediator may vary. For example, in the first study,the levels of TNFα in normal subjects ranged from undetectable (0 pg/ml)to 7.3 pg/ml whereas the levels of IL-1βin normal subjects ranged fromundetectable (0 pg/ml) to 265.5 pg/ml. See, e.g., Nowlan, et al.,Rheumatology 45:31-37, 2006; Mittal & Joshi, J. Indian Rheumatol. Assoc.10:59-60, 2002; Szodoray, et al., Rheumatology 46:417-425, 2007, whichare incorporated herein by reference.

The target value can be a desired ratio of concentrations of two or moreinflammatory mediators in the peripheral blood as determined by a leastsquares fit of the concentration values of the two or more inflammatorymediator. For example, a study assessed the levels of variousinflammatory mediators in subjects with acute graft-versus-host disease(aGVHD) and observed significantly elevated levels of IL-5, IL-6 andIL-10. See, e.g., Fujii, et al., Int. J. Mol. Med. 17:881-885, 2006,which is incorporated herein by reference. The serum level ratios ofIL-5/IL-2, IL-5/IL-4, IL-6/IL-4 were increased in subjects with aGVHD ascompared to transplant subjects with no evidence of aGVHD. In thisinstance, the levels of one or more inflammatory mediators can bealtered to modulate the overall ratio of two or more inflammatorymediators.

Ratios of other inflammatory mediators can be of value in understandingthe types of immune cells activated in association with a diseasecondition. For example, the ratio of Th1/Th2 cytokines may be indicativeof disease severity. Th1 type T-lymphocytes produce IL-2, IL-12, IFNγ,TNFα and TNFβ favoring cell mediated immune responses whereas Th2 typeT-lymphocytes produce IL-4, IL-5, IL-6, IL-10, and IL-13 favoringhumoral responses. The Th2 antibody mediated immune responsepredominates in subjects with sepsis as indicated by a significantlylower Th1/Th2 ratio in subjects with sepsis (median 0.46) as comparedwith non-septic control subjects (median 2.5) and is associated withdecreased resistance to infection (see Andrews & Griffins Brit. J. Nutr.87, Suppl 1:S3-S8, 2002, which is incorporated herein by reference). Theratio of two or more inflammatory mediators can be altered by alteringthe levels of one or more inflammatory mediators.

Intracorporeal Blood Processing

The device for altering the functional structure of one or moreinflammatory mediators in the peripheral blood of a subject having aninflammatory disease or condition can include a device forintracorporeal treatment of the blood. Intracorporeal processing of theperipheral blood of a subject may be accomplished by inserting one ormore devices into one or more intracorporeal locations of a subject. Theintracorporeal device includes one or more sensors for sensing one ormore inflammatory mediators. The intracorporeal device further includesmeans for controllably diverting all or part of the blood flowing inthat part of the subject into one or more treatment chambers. The one ormore treatment chambers can include specific binding agents for bindingone or more specific inflammatory mediators. The one or more treatmentchambers further include one or more reactive components to alter thefunctional structure of one or more inflammatory mediators. Theintracorporeal device further includes a controller that receives senseddata, controls diversion of blood flow, and controls release of the oneor more reactive components for altering the functional structure of oneor more inflammatory mediators.

The intracorporeal device can be inserted into a blood vessel. Theintracorporeal device may be a specialized stent fixed within a specificartery or vein. See, e.g., U.S. Patent Application 2007/0294150 A 1,which is incorporated herein by reference. Alternatively, theintracorporeal device may be any of a number of biocompatible structuresthat may be placed in a blood vessel without impeding blood flow. See,e.g., U.S. Patent Application 2008/0058785 A1, which is incorporatedherein by reference. Alternatively, the intracorporeal device may travelfreely in the circulation as exemplified by a lumen traveling device.See, e.g., U.S. Patent Application 2007/0156211 A1, which isincorporated herein by reference. The device can target to a site ofinflammation in the subject. The device can sense elevated levels ofinflammatory mediators in the peripheral blood or lymphatic system ofthe subject and can target and form a stationary location at, or near, asite of inflammation in the peripheral circulation of the subject. Insome aspects, the intracorporeal device may be incorporated into ashunt, for example, an arteriovenous shunt inserted between an arteryand a vein. Alternatively, the intracorporeal device may be proximal toa blood vessel with controllable access to the blood flow through aconduit. As blood flows through or past the intracorporeal device, oneor more inflammatory mediators may be sensed and the functionalstructure of the inflammatory mediator altered, e.g., antagonizing,inhibiting, binding, blocking, or downregulating the expression of apro-inflammatory mediator, or agonizing, activating, or upregulating theexpression of an anti-inflammatory mediator.

Extracorporeal Blood Processing

The device for altering the functional structure of one or moreinflammatory mediators in the peripheral blood of a subject having aninflammatory disease or condition can include a device providingextracorporeal treatment of the blood. Extracorporeal processing of theperipheral blood of the subject may be accomplished by removing bloodfrom the peripheral circulatory system to an extracorporeal device,altering the functional structure of one or more inflammatory mediators,and returning all or part of the processed blood back to the subject.The extracorporeal device includes one or more sensors for sensing oneor more inflammatory mediators. The extracorporeal device furtherincludes a mechanism for controllably diverting all or part of the bloodflow into one or more treatment chambers. The one or more treatmentchambers can include one or more reactive components, e.g., specificbinding agents, for binding one or more specific inflammatory mediators.The one or more treatment chambers can further include one or morereactive components, e.g. degradative agents, binding agents, or energysources, to alter the functional structure of the one or moreinflammatory mediators. The extracorporeal device can further include acontroller that receives sensed data, controls diversion of blood flow,and controls release of the one or more reactive components to alter thefunctional structure of one or more inflammatory mediators.

Blood may be removed from a subject. In typical high volume dialysistreatment, for example, blood is drawn from the arm through a fistula orgraft between the radial artery and vein. Alternatively, blood may beremoved from a vessel proximal to a critical organ such as for examplethe lungs, heart, kidney, and/or liver. For example, a large-borecentral venous catheter may be used to access fluids in a vein near theheart or just inside the atrium. A Swan-Ganz catheter is a special typeof catheter placed into the pulmonary artery.

Extracorporeal processing of a subject's blood to alter the functionalstructure of one or more inflammatory mediators may be accomplishedusing whole blood. Blood plasma is the liquid component of blood, inwhich the blood cells are suspended. It makes up about 55% of totalblood volume. It is composed of mostly water (90% by volume), andcontains dissolved proteins, glucose, clotting factors, mineral ions,hormones and carbon dioxide (plasma being the main medium for excretoryproduct transportation). Blood plasma is prepared simply by spinning atube of fresh blood in a centrifuge until the blood cells fall to thebottom of the tube. The blood plasma is then poured or drawn off Bloodserum is blood plasma without fibrinogen or the other clotting factors.Alternatively, extracorporeal processing of a subject's blood to alterthe functional structure of one or more inflammatory mediators may beaccomplished using one or more isolated fractions of whole blood. Wholeblood may be fractionated into blood plasma and cellular bloodcomponents by centrifugation and/or membrane filtration using any of anumber of in-line apheresis processes including, but not limited to,plasmapheresis, plasma exchange, plateletpheresis, and leukophoresis.Plasma substantially free of blood cells may be isolated byplasmapheresis in which the plasma is separated from the blood cells bycentrifugation or filtration. The resulting plasma includes a number ofprotein components, including pro-inflammatory and anti-inflammatorymediators. The isolated plasma may be further processed by theextracorporeal device to alter the functional structure of one or moreinflammatory mediators.

The plasma may be further fractionated using a form of hemodialysisand/or hemofiltration through a semi-permeable membrane or filter toinclude and/or exclude components of the plasma based on the relativesize of the components. During hemodialysis, the blood from a subjectflows in a path along one side of a membrane. A dialysate is circulatedon the other side of the membrane and forms a concentration differentialacross the membrane. Liquid and other components carried in the bloodare drawn by the concentration differential across the membrane and outof the blood. During hemofiltration, the blood from a subject flows in apath along a semipermeable membrane, across which a pressure differenceexists. The pores of the membrane have a molecular weight cut-off thatcan pass liquid and components carried in the blood. The device caninclude one or more filters for selective removal of blood components inthe molecular weight range into which most inflammatory mediators fall.See, e.g., U.S. Patent Application 2008/0110830 A1, Tetta, et al.,Kidney Int. Suppl. 63: S69-S71, 2003 which are incorporated herein byreference. Many cytokines, for example, are relatively low molecularweight, electrically neutral proteins, ranging in size from about 8000to about 30,000 daltons. Other inflammatory mediators may range in sizefrom 5,000 to 150,000 daltons. One or more filters with varyingmolecular weight cut-offs may be used to filter the plasma. Whole bloodmay also be used for this purpose. The one or more filtrates containingone or more inflammatory mediators are passed into one or more treatmentchambers of the device to alter the functional structure of the one ormore inflammatory mediators.

Combined Intracorporeal and Extracorporeal Blood Processing

In some aspects, a combination of devices for intracorporeal andextracorporeal processing can be used to sense and/or alter thefunctional structure of one or more inflammatory mediators. For example,one or more intracorporeal devices can be used to sense in real-time thelevels of one or more inflammatory mediators in the peripheral blood ofa subject. Data regarding the levels of one or more inflammatorymediators in the blood of a subject can be transmitted wirelessly to anextracorporeal device that controllably initiates withdrawal of bloodfrom the subject for processing. The extracorporeal device can furthercontrollably initiate release and/or activation of one or more reactivecomponents for altering the functional structure of one or moreinflammatory mediators.

Sensors for Measuring Inflammatory Mediators in the Peripheral Blood

The device includes one or more sensors for qualitatively and/orquantitatively measuring one or more inflammatory mediators, e.g.,pro-inflammatory or anti-inflammatory mediators, in the peripheral bloodof a subject. The one or more sensors can include but are not limited toa biosensor, a chemical sensor, a physical sensor, an optical sensor, ora combination thereof. The one or more sensors can include one or morerecognition elements that recognize one or more inflammatory mediators.The interaction of one or more inflammatory mediators with one or moresensors results in one or more detectable signals. Preferably the one ormore sensors measure in real-time the levels of one or more inflammatorymediators in the peripheral blood of a subject.

The one or more sensors can sense one or more inflammatory mediatorsthat are cytokines including, but not limited to, interferons (IFN)IFN-α, IFN-β, and IFN-γ; interleukins (IL) IL-1, IL-2, IL-3, IL-4, IL-5,IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16,IL-17, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-24, IL-27, IL-28,IL-29, IL-30, IL-31, and IL-32; tumor necrosis factor (TNF) TNF-α andTNF-β; granulocyte colony stimulating factor (G-CSF);granulocyte-macrophage colony stimulating factor (GM-CSF); macrophagecolony-stimulating factor (M-CSF); gelsolin, erythropoietin (EPO); andthrombopoietin (TPO). The one or more inflammatory mediators can be anyof a number of chemotactic cytokines (chemokines) including but notlimited to CC chemokines CCL1 through CCL28 exemplified by RANTES(CCL5), MCP-1 (CCL2), LARC (CCL20), MIP-1 α (CCL3), and MDC (CCL22); CXCchemokines CXCL1 through CXCL17 exemplified by LIX (CXCL5), GCP-2(CXCL6) and BCA-1 (CXCL13); C chemokines XCL1 and XCL2; CX3C chemokineC3CL1 (fractalkine); and chemokine like molecules exemplified by MIF.Other inflammatory mediators include but are not limited toanaphylatoxin fragments C3a, C4a, and C5a from the complement pathway;leukotrienes LTA4, LTB4, LTC4, LTD4, LTE4, and LTF4; prostaglandins;growth factors EGF, FGF-9, FGF-basic, growth hormone, stem cell factor(SCF), TGF-β and VEGF; soluble receptors to tumor necrosis factorreceptor (sTNFr); soluble interleukin receptors sIL-1r and sIL-2r;C-reactive protein; CD11b; histamine; serotonin; apolipoprotein A1;β2-microglobulin; bradykinin; D-dimer; endothelin-1; eotaxin; factorVII; fibrinogen; GST; haptoglobin; IgA; insulin; IP-10; leptin; LIF;lymphotactin; myoglobin; OSM; SGOT; TIMP-1; tissue factor; VCAM-1; VWF;thromboxane; platelet activating factor (PAF); immunoglobulins; andendotoxins such as lipopolysaccharide (LPS); and various exotoxins suchas superantigens, e.g., from, Staphylococcus aureus and Streptococcuspyogenes.

The one or more recognition elements that can identify one or moreinflammatory mediators in the blood can include, but are not limited to,antibodies, antibody fragments, peptides, oligonucleotides, DNA, RNA,aptamers, protein nucleic acids proteins, viruses, enzymes, receptors,bacteria, cells, cell fragments, inorganic molecules, organic molecules,or combinations thereof. The one or more recognition elements can beassociated with one or more substrate integrated into the one or moresensors.

The one or more sensors for sensing one or more inflammatory mediatorscan incorporate one or more recognition elements and one or moremeasurable fluorescent signal. In an embodiment, one or moreinflammatory mediators in the peripheral blood of a subject are capturedby one or more recognition elements and further react with one or morefluorescent second elements. The fluorescence associated with thecaptured one or more inflammatory mediators can be measured usingfluorescence spectroscopy. Alternatively, the fluorescence signal can bedetected using at least one charged-coupled device (CCD) and/or at leastone complimentary metal-oxide semiconductor (CMOS).

In an aspect, the one or more sensors can use Förster or fluorescenceresonance energy transfer (FRET) to sense one or more inflammatorymediators in the peripheral blood of a subject. FRET is adistance-dependent interaction between the electronic excited states oftwo dye molecules in which excitation is transferred from a donormolecule to an acceptor molecule without emission of a photon. In someaspects, interaction of a donor molecule with an acceptor molecule maylead to a shift in the emission wavelength associated with excitation ofthe acceptor molecule. In other aspects, interaction of a donor moleculewith an acceptor molecule may lead to quenching of the donor emission.The one or more recognition elements associated with the one or moresensors may include at least one donor molecule and at least oneacceptor molecule. Binding of one or more inflammatory mediators to therecognition element may result in a conformation change in therecognition element, leading to changes in the distance between thedonor and acceptor molecules and changes in measurable fluorescence. Therecognition element may be a cell, an antibody, an aptamer, a receptoror any other molecule that changes conformation or signaling in responseto binding one or more inflammatory mediators.

A variety of donor and acceptor fluorophore pairs may be considered forFRET associated with the recognition element including, but not limitedto, fluorescein and tetramethylrhodamine; IAEDANS and fluorescein;fluorescein and fluorescein; and BODIPY FL and BODIPY FL. A number ofAlexa Fluor (AF) fluorophores (Molecular Probes-Invitrogen, Carlsbad,Calif., USA) may be paired with other AF fluorophores for use in FRET.Some examples include, but are not limited, to AF 350 with AF 488; AF488 with AF 546, AF 555, AF 568, or AF 647; AF 546 with AF 568, AF 594,or AF 647; AF 555 with AF594 or AF647; AF 568 with AF6456; and AF594with AF 647.

The cyanine dyes Cy3, Cy5, Cy5.5 and Cy7, which emit in the red and farred wavelength range (>550 nm), offer a number of advantages forFRET-based detection systems. Their emission range is such thatbackground fluorescence is often reduced and relatively large distances(>100 Å) can be measured as a result of the high extinction coefficientsand good quantum yields. For example, Cy3, which emits maximally at 570nm and Cy5, which emits at 670 nm, may be used as a donor-acceptor pair.When the Cy3 and Cy5 are not proximal to one another, excitation at 540nm results only in the emission of light by Cy3 at 590 nm. In contrast,when Cy3 and Cy5 are brought into proximity by a conformation change inan aptamer, antibody, or receptor, for example, excitation at 540 nmresults in an emission at 680 nm. Semiconductor quantum dots (QDs) withvarious excitation/emission wavelength properties may also be used togenerate a fluorescence based sensor.

Quenching dyes may be used as part of the binder element to quench thefluorescence of visible light-excited fluorophores. Examples include,but are not limited, to DABCYL, the non-fluorescing diarylrhodaminederivative dyes QSY 7, QSY 9 and QSY 21 (Molecular Probes, Carlsbad,Calif., USA), the non-fluorescing Black Hole Quenchers BHQ0, BHQ1, BHQ2,and BHQ3 (Biosearch Technologies, Inc., Novato, Calif., USA) and Eclipse(Applera Corp., Norwalk, Conn., USA). A variety of donor fluorophore andquencher pairs may be considered for FRET associated with therecognition element including, but not limited to, fluorescein withDABCYL; EDANS with DABCYL; or fluorescein with QSY 7 and QSY 9. Ingeneral, QSY 7 and QSY 9 dyes efficiently quench the fluorescenceemission of donor dyes including blue-fluorescent coumarins, green- ororange-fluorescent dyes, and conjugates of the Texas Red and Alexa Fluor594 dyes. QSY 21 dye efficiently quenches all red-fluorescent dyes. Anumber of the Alexa Fluor (AF) fluorophores (MolecularProbes-Invitrogen, Carlsbad, Calif., USA) may be paired with quenchingmolecules as follows: AF 350 with QSY 35 or DABCYL; AF 488 with QSY 35,DABCYL, QSY7 or QSY9; AF 546 with QSY 35, DABCYL, QSY7 or QSY9; AF 555with QSY7 or QSY9; AF 568 with QSY7, QSY9 or QSY21; AF 594 with QSY21;and AF 647 with QSY 21.

The one or more sensor for sensing one or more inflammatory mediatorscan use the technique of surface plasmon resonance (for planar surfaces)or localized surface plasmon resonance (for nanoparticles). Surfaceplasmon resonance involves detecting changes in the refractive index ona sensor surface in response to changes in molecules bound on the sensorsurface. The surface of the sensor may be a glass support or other solidsupport coated with a thin film of metal, for example, gold. The sensorsurface may further carry a matrix to which is immobilized one or morerecognition elements that recognize one or more inflammatory mediators.The one or more recognition elements that recognize one or moreinflammatory mediators may be antibodies or fragments thereof,oligonucleotide or peptide based aptamers, receptors of inflammatorymediators or fragments thereof, artificial binding substrates formed bymolecular imprinting, or any other examples of molecules and orsubstrates that bind inflammatory mediators. As blood or bloodcomponents from the subject passes by the sensor surface, one or moreinflammatory mediators may interact with one or more recognitionelements on the sensor surface. The sensor is illuminated bymonochromatic light. Resonance occurs at a specific angle of incidentlight. The resonance angle depends on the refractive index in thevicinity of the surface, which is dependent upon the concentration ofmolecules on the surface. An example of instrumentation that usessurface plasmon resonance is the BIACORE system (Biacore, Inc.—GEHealthcare, Piscataway, N.J.) which includes a sensor microchip, a laserlight source emitting polarized light, an automated fluid handlingsystem, and a diode array position sensitive detector. See, e.g.,Raghavan & Bjorkman Structure 3:331-333, 1995, which is incorporatedherein by reference.

The one or more sensors can be one or more label-free optical biosensorsthat incorporate other optical methodologies, e.g., interferometers,waveguides, fiber gratings, ring resonators, and photonic crystals. See,e.g., Fan, et al., Anal. Chim. Acta 620:8-26, 2008, which isincorporated herein by reference. For example, reflectometricinterference spectroscopy can be used to monitor in real-time theinteraction of the inflammatory mediator interferon 2 with ananti-interferon 2 antibody. See, e.g., Piehler & Schreiber, Anal.Biochem. 289:173-186, 2001, which is incorporated herein by reference.

The one or more sensors for sensing one or more inflammatory mediatorscan be one or more microcantilevers. A microcantilever can act as abiological sensor by detecting changes in cantilever bending orvibrational frequency in response to binding of one or more inflammatorymediators to the surface of the sensor. In an aspect the sensor can bebound to a microcantilever or a microbead as in an immunoaffinitybinding array. In another aspect, a biochip can be formed that usesmicrocantilever bi-material formed from gold and silicon, as sensingelements. See, e.g. Vashist J. Nanotech Online 3:DO:10.2240/azojono0115, 2007, which is incorporated herein by reference.The gold component of the microcantilever can be coated with one or morerecognition elements which upon binding one or more inflammatorymediators causes the microcantilever to deflect. Aptamers or antibodiesspecific for one or more inflammatory mediators can be used to coatmicrocantilevers. See, e.g., U.S. Pat. No. 7,097,662, which isincorporated herein by reference. The one or more sensor can incorporateone or more methods for microcantilever deflection detection including,but not limited to, piezoresistive deflection detection, opticaldeflection detection, capacitive deflection detection, interferometrydeflection detection, optical diffraction grating deflection detection,and charge coupled device detection. In some aspects, the one or moremicrocantilever can be a nanocantilever with nanoscale components. Theone or more microcantilevers and/or nanocantilevers can be arranged intoarrays for detection of one or more inflammatory mediators. Bothmicrocantilevers and nanocantilevers can find utility inmicroelectomechnical systems (MEMS) and/or nanoelectomechnical systems(NEMS) associated with an extracorporeal or intracorporeal device.

The one or more sensor for sensing one or more inflammatory mediator canbe a field effect transistor (FET) based biosensor. In this aspect, achange in electrical signal is used to detect interaction of one or moreanalytes with one or more components of the sensor. See, e.g., U.S. Pat.No. 7,303,875, which is incorporated herein by reference.

The one or more sensors for sensing one or more inflammatory mediatorscan incorporate electrochemical impedance spectroscopy. Electrochemicalimpedance spectroscopy can be used to measure impedance across a naturaland/or artificial lipid bilayer. The sensor can incorporate anartificial bilayer that is tethered to the surface of a solid electrode.One or more receptor can be embedded into the lipid bilayer. The one ormore receptors can be ion channels that open and close in response tobinding of a specific analyte. The open and closed states can bequantitatively measured as changes in impedance across the lipidbilayer. See, e.g., Yang, et al., IEEE SENSORS 2006, EXCO, Daegu,Korea/Oct. 22-25, 2006, which is incorporated herein by reference.

The one or more sensors for sensing one or more inflammatory mediatorcan be cells that include one or more binding elements which when boundto one or more inflammatory mediator induces a measurable or detectablechange in the cells. The cells may emit a fluorescent signal in responseto interacting with one or more inflammatory mediators. For example, abioluminescent bioreporter integrated circuit may be used in whichbinding of a ligand to a cell induces expression of reporter polypeptidelinked to a luminescent response (U.S. Pat. No. 6,673,596, [Durick &Negulescu Biosens. Bioelectron. 16:587-592, 2001] which are incorporatedherein by reference. Alternatively, the one or more cell may emit anelectrical signal in response to interacting with one or moreinflammatory mediator. In a further aspect, an implantable biosensor maybe used which is composed of genetically-modified cells that respondedto ligand binding by emitting a measurable electrical signal. See U.S.Patent Application 2006/0234369 A1; which are incorporated herein byreference.

The device can further include one or more sensors for sensing one ormore physiological parameters in the subject. Examples of physiologicalparameters include but are not limited to body temperature, respirationrate, pulse, blood pressure, edema, oxygen saturation, pathogen levels,or toxin levels.

Controller In Communication With and Responsive to a Sensor

The device can further include a controller that is in communicationwith and configured to be informed by the one or more sensors. The oneor more sensors can transmit data to the controller regarding thedetection or levels (relative or absolute) of one or more inflammatorymediators in the peripheral blood of a subject. The controller can beintegrated to the extracorporeal or intracorporeal device.Alternatively, the controller can be a separate component of the devicethat receives and transmits data and/or commands either with or withoutwires. For example, an intracorporeal device can send data regarding thesensed levels of one or more inflammatory mediators to an externalcontroller through a wireless signal.

The controller can compare the input data regarding the one or moreinflammatory mediators in the blood of a subject with stored data. Thecontroller itself can include the stored data. Alternatively, thecontroller can have access to one or more remote databases that includethe stored data. The stored data may be data regarding the normal levelof one or more inflammatory mediators in normal or healthy subjectswithout inflammatory conditions. The stored data may further includedata regarding the baseline level of one or more inflammatory mediatorsin a subject prior to an inflammatory condition. The stored data mayfurther include data regarding the level of one or more inflammatorymediators in a subject at one or more previous time points. Thecontroller assesses the most recently obtained input data with thestored data and is configured to controllably initiate steps to alterthe functional structure of one or more inflammatory mediators in theperipheral blood of a subject.

In response to input data, the controller can cause the device tocontrollably divert all or part of the blood of a subject into one ormore treatment chambers. Access to one or more treatment chambers can becontrolled by a flow-modulating element. A flow-modulating element maybe a gate, a valve, a louver, a splitter or flow divider, a filter, abaffle, a channel restriction, a retractable iris, or other structurethat controllably limits access of the blood flow to a treatmentchamber. The controller can send a signal to the flow-modulating elementindicating whether or not all or part of the flow of blood should bediverted into a treatment chamber.

The controller can further controllably initiate release or activationof one or more reactive components designed to alter the functionalstructure of one or more inflammatory mediators. The one or morereactive components can be controllably released or activated by thecontroller in the one or more treatment chambers of the device. In anaspect, the controller can release of one or more modulator into theperipheral blood of a subject to modulate the activity and/or expressionof one or more inflammatory mediators. Alternatively, the controller cansend data regarding the levels of one or more inflammatory mediators inthe peripheral blood of a subject to the subject, to one or more thirdparty individuals such as a physician or other caregiver, to a computingdevice, or to a combination thereof. The subject and/or caregiver orcomputing device can choose to initiate steps to alter the functionalstructure of one or more inflammatory mediators, to release modulatorsinto the circulation, or a combination thereof.

The controller can also include one or more algorithms that providecomputational models of inflammation. A computational model ofinflammation may include information regarding, for example, a varietyof interrelated signaling pathways involved in pro-inflammatory andanti-inflammatory processes. The computational model may further informdecisions made by the controller. See, e.g., U.S. Pat. No. 7,415,359 B2;U.S. Patent Applications 2007/0083333 A1, 2008/0201122 A1; Vodovotz, etal., Curr. Opin. Crit. Care. 10:383-390, 2004; Zenker, et al., PLoSComput. Biol. 3(11):e204, 2007; Li, et al., PLoS ONE 3(7):e2789, 2008;Vodovotz, et al., PLoS Comput. Biol. 4:e1000014, 2008; An, TheoreticalBiology Medical Modeling 5:11, 2008, each of which is incorporatedherein by reference.

Device Including One or More Reactive Components

A device is disclosed that includes a means for modulating aphysiological effect of one or more inflammatory mediators in theperipheral blood of a subject. The means for modulating thephysiological effect can include, for example, one or more reactivecomponents that are used to alter the functional structure of one ormore inflammatory mediators in the peripheral blood of the subject. In afurther aspect, the one or more reactive components can be used to alterthe functional structure of one or more elements of one or moreinflammatory signaling pathway. A reactive component includes, but isnot limited to, a denaturing agent, a degradative agent, a bindingagent, an energy source, or a combination thereof. A reactive componentcan further include a modulator that modulates the activity of one ormore inflammatory mediators. The one or more reactive components areincorporated into or released within one or more treatment chambersassociated with the device. Alternatively, the one or more reactivecomponents are diffusible components released from a reservoir of thedevice into the peripheral blood of the subject.

The device includes one or more reactive component that can modulate thephysiological effect of one or more inflammatory mediators, e.g., alterthe functional structure of the one or more inflammatory mediators. Theone or more inflammatory mediators can be cytokines, including but notlimited to, interferons (IFN) IFN-α, IFN-β, and IFN-γ; interleukins (IL)IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11,IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, IL-21,IL-22, IL-23, IL-24, IL-27, IL-28, IL-29, IL-30, IL-31, and IL-32; tumornecrosis factor (TNF) TNF-α and TNF-β; granulocyte colony stimulatingfactor (G-CSF); granulocyte-macrophage colony stimulating factor(GM-CSF); macrophage colony-stimulating factor (M-CSF); erythropoietin(EPO); and thrombopoietin (TPO). The one or more inflammatory mediatorscan be any of a number of chemotactic cytokines (chemokines) including,but not limited to, CC chemokines CCL1 through CCL28 exemplified byRANTES (CCL5), MCP-1 (CCL2), LARC(CCL20), MIP-1 α (CCL3), and MDC(CCL22); CXC chemokines CXCL1 through CXCL17 exemplified by LIX (CXCL5),GCP-2 (CXCL6) and BCA-1 (CXCL13); C chemokines XCL1 and XCL2; CX3Cchemokine C3CL1 (fractalkine); and chemokine like molecules exemplifiedby MIF. Other inflammatory mediators include but are not limited toanaphylatoxin fragments C3a, C4a, and C5a from the complement pathway;leukotrienes LTA4, LTB4, LTC4, LTD4, LTE4, and LTF4; prostaglandins;growth factors EGF, FGF-9, FGF-basic, growth hormone, stem cell factor(SCF), TGF-β and VEGF; soluble receptors to tumor necrosis factorreceptor (sTNFr); soluble interleukin receptors sIL-1r and sIL-2r;C-reactive protein; CD11b; histamine; serotonin; apolipoprotein A1;β2-microglobulin; bradykinin; D-dimer; endothelin-1; eotaxin; factorVII; fibrinogen; GST; haptoglobin; IgA; insulin; IP-10; leptin; LIF;lymphotactin; myoglobin; OSM; SGOT; TIMP-1; tissue factor; VCAM-1; VWF;thromboxane; platelet activating factor (PAF); immunoglobulins; andendotoxins such as lipopolysaccharide (LPS); and various exotoxins suchas superantigens, e.g., from Staphylococcus aureus and Streptococcuspyogenes.

In some aspects, the device including the one or more reactivecomponents can modulate the physiological effect, e.g., alter thefunctional structure, of one or more elements of one or moreinflammatory signaling pathways. For example, activation of Toll-likereceptors (TLRs) or IL-1 receptor can induce inflammation in immunecells via shared signaling cascades. TLRs are expressed in or onmonocytes, macrophages, dendritic cells and microglia. TLRs recognizeand respond to pathogen-associated molecular patterns (PAMPs) such aslipopolysaccharide, lipoteichoic acid, DNA with non-methylatedcytosine-guanine motifs, zymosan, and/or viral double-stranded RNA. TheTLR family members and the IL-1 receptor have a unique intracellularToll/IL-1 receptor signaling domain which in response to activation,transduces the signal to a family of IL-1 receptor-associated kinases(IRAK). Phosphorylation of IRAK induces cascades of signaling throughtumor necrosis factor receptor-associated factor 6 which, in turn,transduces the signal to IκB kinase-β and to mitogen-activated proteinkinase. This signaling results in transcriptional responses, mediatedprimarily by nuclear factor-κB, extracellular-signal regulated kinaseand stress-activated protein kinases, such as c-Jun N-terminal kinase(JNK) and p38, leading to expression of proinflammatory cytokines. Oneor more reactive components of the device can modulate the activity ofone or more elements of this or other inflammatory signaling pathways.Extensive examples of signaling pathways associated with inflammationand other cellular processes may be accessed in the scientificliterature and/or through a database, for example, Database of CellSignaling (see, e.g., Goeddel & Chen, Tumor Necrosis Factor Pathway.Sci. Signal. Connections Map in the Database of Cell Signaling, as seen6 Nov. 2008; CMP_(—)7107; “TNF-R1 Signaling: A Beautiful Pathway” Chen &Goeddel Science 296: 1634-1635, 2002) and/or the UCSD-Nature SignalingGateway (see, e.g., Nat. Cell Biol. 6: 1, 2004; each of which isincorporated herein by reference.

Binding agents remove one or more inflammatory mediators from theperipheral blood. The device can include one or more reactive componentsthat are binding agents designed to remove one or more inflammatorymediators from the peripheral blood of a subject. The one or morebinding agents can be used alone to selectively or non-selectivelysequester one or more inflammatory mediators. Alternatively, the one ormore binding agents can be used to capture one or more inflammatorymediators in combination with treatment including one or more additionalreactive components, e.g., a denaturing agent, a degradative agent, amodulator, an energy source, or a combination thereof. Following bindingof the one or more inflammatory mediators to the one or more bindingagents in a treatment chamber, one or more additional reactivecomponents can be provided to alter the functional structure of the oneor more inflammatory mediators.

The one or more binding agents can include absorbent material thatnon-selectively binds one or more inflammatory mediators. The absorbentmaterial may include, but is not limited to, silica, activated charcoal,nonionic or uncharged resins or polymers, ionic or charged resins orpolymers, anion exchange resins or polymers, cation exchange resins orpolymers, neutral exchange resins or polymers, immobilized polymyxin B,immobilized monoclonal antibodies, immobilized inflammatory mediatorreceptors, immobilized specific antagonists, cellulose, cellulosederivatives, synthetic materials, polysulfone, polyacrylonitrile,polymethylmethacrylate, polyvinyl-alcohol, polyamide, polycarbonate,polystyrene-derivative fibers, and any combination thereof. Specificexamples of absorbent materials that have been used in animal andclinical studies for non-specific binding of inflammatory mediatorsinclude, but are not limited to, polystyrene-divinylbenzene copolymerbeads with biocompatible polyvinylpyrrolidone coating (CYTOSORB,MedaSorb Corporation, NJ, USA) and 2-methacryloyloxyethylphosphorylcholine (MPCF-X; see, e.g., Nakada, et al., Transfus. Apher.Sci. 35:253-264, 2006, which is incorporated herein by reference.

The one or more binding agents can selectively bind one or moreinflammatory mediators. A selective binding agent of one or moreinflammatory mediators can include, but is not limited to, an antibodyor fragments thereof, an oligonucleotide or peptide based aptamer, aninflammatory mediator receptor or parts thereof, an artificial bindingsubstrate formed by molecular imprinting, or other examples ofbiomolecules and or substrates that bind inflammatory mediators.

The one or more binding agents can include one or more antibodies thatbind one or more inflammatory mediators. Antibodies or fragments thereoffor use as one or more binding agents of inflammatory mediators mayinclude, but are not limited to, monoclonal antibodies, polyclonalantibodies, Fab fragments of monoclonal antibodies, Fab fragments ofpolyclonal antibodies, Fab₂ fragments of monoclonal antibodies, and Fab₂fragments of polyclonal antibodies, chimeric antibodies, non-humanantibodies, fully human antibodies, among others. Single chain ormultiple chain antigen-recognition sites can be used. Multiple chainantigen-recognition sites can be fused or unfused. Antibodies orfragments thereof may be generated using standard methods. See, e.g.,Harlow & Lane (Antibodies: A Laboratory Manual, Cold Spring HarborLaboratory Press; 1^(st) edition 1988), which is incorporated herein byreference. Alternatively, an antibody or fragment thereof directedagainst one or more inflammatory mediators may be generated, forexample, using phage display technology. See, e.g., Kupper, et al. BMCBiotechnology 5:4, 2005, which is incorporated herein by reference. Anantibody, a fragment thereof, or an artificial antibody, e.g., Affibody®artificial antibodies (Affibody A B, Bromma, Sweden) can be preparedusing in silico design (Knappik et al., J. Mol. Biol. 296: 57-86, 2000,which is incorporated herein by reference. In some aspects, antibodiesdirected against one or more inflammatory mediators may be availablefrom a commercial source (from, e.g., Novus Biological, Littleton,Colo.; Sigma-Aldrich, St. Louis, Mo.; United States Biological,Swampscott, Mass.).

The one or more binding agents can include one or more aptamers thatbind one or more inflammatory mediators. The aptamer can be anoligonucleotide RNA- or DNA-based aptamer. Aptamers are artificialoligonucleotides (DNA or RNA) that can bind to a wide variety ofentities (e.g., metal ions, small organic molecules, proteins, andcells) with high selectivity, specificity, and affinity. Aptamers may beisolated from a large library of 10¹⁴ to 10¹⁵ random oligonucleotidesequences using an iterative in vitro selection procedure often termed“systematic evolution of ligands by exponential enrichment” (SELEX).See, e.g., Cao, et al., Current Proteomics 2:31-40, 2005; Proske, etal., Appl. Microbiol. Biotechnol. 69:367-374, 2005; Jayasena Clin. Chem.45:1628-1650, 1999, which are incorporated herein by reference. Ingeneral, SELEX may be used to generate aptamers against inflammatorymediators, for example, cytokines and growth factors. See, e.g.,Guthrie, et al., Methods 38:324-330, 2006, which is incorporated hereinby reference. In an aspect, SELEX may be used to generate an RNA aptameragainst the inflammatory mediator TNF-α. See, e.g., U.S. Pat. No.7,309,789, which is incorporated herein by reference.

In an aspect, the one or more binding agents can include one or moreaptamers that are peptide based aptamers. Peptide aptamers areartificial proteins in which inserted peptides are expressed as part ofthe primary sequence of a structurally stable protein. See, e.g.,Crawford, et al., Brief. Funct. Genomic Proteomic 2:72-79, 2003, whichis incorporated herein by reference. Peptide aptamers may be generatedby screening an inflammatory mediator against yeast two-hybridlibraries, yeast expression libraries, bacterial expression librariesand/or retroviral libraries. Peptide aptamers may have bindingaffinities comparable to antibodies.

In a further aspect, the one or more binding agents can include one ormore novel peptides. Novel peptides that bind selective targets may begenerated, for example, using phage display methodologies. See, e.g.,Spear, et al., Cancer Gene Ther. 8:506-511, 2001, which is incorporatedherein by reference. In this aspect, the phage express novel peptides onthe surface as fusion proteins in association with a phage major orminor coat protein and may be screened for binding interaction with oneor more inflammatory mediators.

The one or more binding agents can include one or more inflammatorymediator receptors that bind one or more inflammatory mediators. All orpart of an inflammatory mediator receptor may be used as a specificbinding agent. Examples of inflammatory mediator receptors include, butare not limited to, type I cytokine receptors such as type 1 interleukinreceptors, erythropoietin receptor, GM-CSF receptor, G-CSF receptor,growth hormone receptor, oncostatin M receptor, leukemia inhibitoryfactor receptor; type II cytokine receptors such as type II interleukinreceptors, interferon-α/β receptors, interferon-γ receptor; many membersof the immunoglobulin superfamily such as interleukin-1 receptor, CSF1,c-kit receptor, interleukin-18 receptor; tumor necrosis factor (TNF)receptor family such as TNF receptor 1 (TNF-R1), TNF receptor 2(TNF-R2), CD27, CD40, and lymphotoxin β receptor; chemokine receptorsincluding serpentine CCR and CXCR receptors, such as CCR1 and CXCR4, andinterleukin-8 receptor; TGF β receptors such as TGF β receptor 1 and TGFβ receptor 2. See Ozaki and Leonard, J. Biol. Chem. 277:29355-29358,2002, which is incorporated herein by reference

The one or more binding agents can include one or more artificialbinding substrates for one or more inflammatory mediators formed by theprocess of molecular imprinting. In the process of molecular imprinting,a template is combined with functional monomers which upon cross-linkingform a polymer matrix that surrounds the template. See Alexander, etal., J. Mol. Recognit. 19:106-180, 2006, which is incorporated herein byreference. Removal of the template leaves a stable cavity in the polymermatrix that is complementary in size and shape to the template. In anaspect, functional monomers of acrylamide and ethylene glycoldimethacrylate may be mixed with one or more inflammatory mediators inthe presence of a photoinitiator and ultraviolet irradiation used tocross-link the monomers. The resulting polymer may be crushed or groundinto smaller pieces and washed to remove the one or more inflammatorymediators, leaving a particulate matrix material capable of binding oneor more inflammatory mediators. Examples of other functional monomers,cross-linkers and initiators may be used to generate an artificialbinding substrate are provided. See, e.g., U.S. Pat. No. 7,319,038;Alexander, et al., J. Mol. Recognit. 19:106-180, 2006, which areincorporated herein by reference. In a further aspect, hydrogels may beused for molecular imprinting. See, e.g., Byrne et al., “Molecularimprinting within hydrogels”, Advanced Drug Delivery Reviews, 54:149-161, 2002, which is incorporated herein by reference. Other examplesof synthetic binders are provided. See, e.g., U.S. Pat. Nos. 6,255,461;5,804,563; 6,797,522; 6,670,427; and 5,831,012; and U.S. PatentApplication 20040018508; and Ye and Haupt, Anal Bioanal Chem. 378:1887-1897, 2004; Peppas and Huang, Pharm Res. 19: 578-587 2002, whichare incorporated herein by reference.

Reactive components can include denaturing agents that alter thefunctional structure of one or more inflammatory mediators. The deviceincluding one or more reactive components can include one or moredenaturing agents. The functional structure of one or more inflammatorymediators can be altered by the process of denaturation in which thesecondary, tertiary or quaternary structure of one or more inflammatorymediators are altered by denaturing agents. Examples of denaturingagents include, but are not limited to, acids such as acetic acid,trichloroacetic acid (TCA), sulfosalicyclic acid, picric acid; solventssuch as methanol, ethanol, and acetone; cross-linking agents such asformaldehyde and gluteraldehyde; chaotropic agents such as urea,guanidinium chloride, and lithium perchlorate; and disulfide bondreducers such as 2-mercaptoethanol, dithithreitol, and TCEP. In anaspect, acids may be used to denature a protein molecule by exposing theprotein molecule to a pH below its isoelectric point. Under theseconditions, the protein molecule will lose its negative charge andretain only positive charges. The like positive charges may repel oneanother and in areas of large charge density, the intramolecularrepulsion may be sufficient enough to cause unfolding of the protein.The one or more denaturing agents may be incorporated into or releasedwithin one or more treatment chambers of the device. Alternatively, theone or more denaturing agents may be released by the device asdiffusible agents into the peripheral blood.

Reactive components can include degradative agents that alter thefunctional structure of one or more inflammatory mediators. Thefunctional structure of one or more inflammatory mediators can bealtered by the one or more degradative agents that act by breakingpeptide bonds within the primary amino acid sequence of the one or moreinflammatory mediators. The one or more degradative agents can includeany of a number of agents designed to cleave one or more peptide bondsof the primary amino acid sequence of one or more inflammatorymediators. Examples of degradative agents, include but are not limitedto proteases, strong acids, strong bases, free radicals, natural orsynthetic proteasomes, or photoactivatable agents. The one or moredegradative agents may be incorporated into or released within one ormore treatment chambers of the device. Alternatively, the one or moredegradative agents may be released by the device as diffusible agentsinto the peripheral blood.

The device including one or more degradative agents can include one ormore proteases. Examples of proteases include, but are not limited to,serine proteases, e.g., as trypsin, chymotrypsin, elastase, dipeptidylpeptidase IV, and subtilisin; cysteine proteases, e.g., papain,cathepsins, caspases, calpains; aspartic acid proteases, e.g., pepsin,renin, and HIV-proteases; metalloproteases, e.g. carboxypeptidases,aminopeptidases, and matrix metalloproteases, e.g. MMP1 through MMP28.The one or more proteases may be free in solution. Alternatively, theone or more proteases may be bound to a substrate. In an aspect, trypsinmay be bound to glass beads. See, e.g., Lee, et al., J. Dairy Sci.,58:473-476, 1974, which is incorporated herein by reference.Alternatively, trypsin and other proteases may be bound to an agarosematrix. Sources of immobilized proteases including trypsin and pepsinare available from commercial sources (Pierce Chemicals, Rockford, Ill.;Applied Biosystems, Foster City, Calif.).

The device including one or more degradative agents can include anatural or synthetic complex of proteases. In an aspect, the one or moreinflammatory mediators may be subject to degradation using proteasomes.A proteasome is a naturally occurring large protein complex thatcontains multiple subunits. The complex includes several proteaseactivities, for example, chymotrypsin-like activity, trypsin-likeactivity, glutamic acid protease activity, and threonine proteaseactivity. Proteasome complexes may be purified from fractionated cellsusing ultracentrifugation through a 10-40% glycerol gradient. See, e.g.,Pervan, et al., Mol. Cancer. Res. 3:381-390, 2005, which is incorporatedherein by reference. Proteasomes may be isolated using a commerciallyavailable isolation kit. (Proteasome Isolation Kit, Human 539176-1KIT,Calbiochem (EMD Chemicals, Inc.; Gibbstown, N.J.).

The device including one or more degradative agents can include an agentthat selectively targets one or more inflammatory mediators fordegradation. In an aspect, the one or more inflammatory mediators may becovalently tagged with ubiquitin for selective destruction byproteasomes. Ubiquitin is a small and highly conserved protein. Anisopeptide bond links the terminal carboxyl of ubiquitin to the ∈-aminogroup of a lysine residue of a protein targeted for degradation. Thejoining of ubiquitin to the targeted protein is ATP-dependent. Threeenzymes are involved, designated E1, E2 and E3. Initially, the terminalcarboxyl group of ubiquitin is joined in an ATP-dependent thioester bondto a cysteine residue on ubiquitin-activating enzyme (E1). The ubiquitinis then transferred to a sulfhydryl group on a ubiquitin-conjugatingenzyme (E2). A ubiquitin-protein ligase (E3) then promotes transfer ofubiquitin from E2 to the ∈-amino group of a lysine residue of a proteinrecognized by that E3, forming an isopeptide bond. There are distinctubiquitin ligases with differing substrate specificity. In addition,some proteins have specific sequences termed a “destruction box” that isa domain recognized by a corresponding ubiquitin ligase. In general, E1,E2, and E3 may be isolated from natural sources or generated usingstandard molecular biology techniques and used to ubiquinate proteins invitro. See, e.g., Chen, et al., EMBO Rep. 2:933-938, 2001, which isincorporated herein by reference. In some aspects, the E2 ligase may begenetically engineered in such a manner as to recognize a specificsubstrate. See, e.g., Colas, et al., PNAS 97:13720-13725, 2000, which isincorporated herein by reference. The device including the treatmentchamber may further include one or more genetically engineered E2 ligaseenzymes capable of adding ubiquitin to and facilitating degradation ofthe one or more inflammatory mediators in the peripheral blood of thesubject.

In a further aspect, the ubiquitin may be indirectly associated with theone or more targeted inflammatory mediators. In an aspect, the ubiquitinmay be linked to an antibody or an aptamer that specifically binds oneor more inflammatory mediators. Binding of the ubiquitin-labeledantibody or aptamer to one or more inflammatory mediators may mark theprotein conjugate for degradation by proteasomes.

The device including one or more degradative agents can include a strongacid. Acid hydrolysis may result in degradation of the one or moreinflammatory mediators. In this aspect, strong acids such ashydrochloric acid or sulfuric acid may be used to break thecarbon-nitrogen peptide bond. Degradation of one or more inflammatorymediators by acid hydrolysis may be optionally performed in combinationwith elevated temperature, a nitrogen atmosphere and or microwaveenergy.

The device including one or more degradative agents can include one ormore free radical reactive oxygen species. Examples of reactive oxygenspecies include, but are not limited to, singlet molecular oxygen,superoxide ion, hydrogen peroxide, hypochlorite ion, hydroxyl radical.Reactive oxygen species can react directly with proteins, targetingpeptide bonds or amino acid side chains. See, e.g., Davies, Biochem.Biophys. Res. Commun. 305:761-770, 2003, which is incorporated herein byreference. A number of the reactions mediated by reactive oxygen specieslead to introduction of carbonyl groups into the protein which in turnmay result in inactivation of the protein by cleavage of the peptidebound to yield lower-molecular weight products, cross-linking ofproteins to yield higher-molecular weight products, or loss of catalyticfunction or structural function by distorting secondary and tertiarystructure, or combination thereof. Reactive oxygen species may induce aamidation, diamide, glutamate oxidation and or proline oxidation whichcan lead to cleavage of peptide bonds. Reactive oxygen species may beformed by the interaction of biological molecules with componentsincluding, but not limited to, ionizing radiation, as a byproduct ofcellular respiration, and dedicated enzymes such as NADPH oxidase andmyeloperoxidase.

In an aspect, the device including one or more degradative agents caninclude reactive oxygen species that are singlet oxygen species. Singletoxygen can cause damage to both the side-chains and backbone of aminoacids, peptides, and proteins. See, e.g., Davies, Biochem. Biophys. Res.Commun. 305:761-770, 2003, which is incorporated herein by reference.Singlet oxygen species may react with tryptophan, tyrosine, histidine,methionine and or cysteine and cystine residues within a polypeptide andmay cause increased susceptibility to proteolytic enzymes, an increasedextent/susceptibility to unfolding, changes in conformation, an increasein hydrophobicity, and changes in binding of co-factor and metal ions.In particular, the interaction of tyrosine with singlet oxygen speciesmay lead to fragmentation or cleavage of the polypeptide. See, e.g.,Davies, Biochem. Biophys. Res. Commun. 305:761-770, 2003, which isincorporated herein by reference.

The device including one or more degradative agents can include one ormore singlet oxygen species generated by a photosensitizer, a chemicalwhich upon exposure to a given wavelength of light emits singlet oxygenspecies. Examples of photosensitizers include, but are not limited to,porphyrin derivatives such as Photofin, which is excited by red light at630 nm; chlorins and bacteriochlorins such as bonellin (maximumabsorbance 625 nm), mono-L-aspartyl chlorine e6 (max abs 654),m-tetrahydroxyphenyl chlorine (mTHPC, max abs 652 nm), and tinetiopurpurin (SnET2, maximum absorbance 660 nm); benzoporphyrinderivatives such as veteroporfin (also labeled BPD-MA, maximumabsorbance 690 nm), 5-aminolaevulinic acid (ALA, porphoryin precursor toPpIX (maximum absorbance 635 nm)); texaphyrins such as lutetiumtexaphyrin (Lu-Tex, maximum absorbance 732), Phthalocyanines andnaphthalocyanines (maximum absorbance 670-780 nm); and cationicphotosensitizers such as rhodamine 123 and methylene blue. See, e.g.,Prasad (2003) Introduction to Biophotonics, John Wiley & Sons, Inc.Hoboken, N.J. Tunable quantum dots (QDs), especially those absorbing inthe wavelength range of 600 to 800 nm, also emit singlet oxygen speciesin response to light and may be useful as photosensitizers. See, e.g.,Samia, et al. (2006) Photochem. Photobiol. 82:617-625, which isincorporated herein by reference.

Modulators can alter the functional structure of one or moreinflammatory mediators. The device can include one or more reactivecomponents that are one or more modulators that either directly orindirectly modulate the activity of one or more inflammatory mediatorsin the peripheral blood of a subject. The one or more modulators can beincorporated into or released within one or more treatment chambers ofthe device. Alternatively, the one or more modulators can be released bythe device as diffusible agents into the peripheral blood. A modulatormay alter, modify, reduce or eliminate the activity of one or moreinflammatory mediators by preventing the binding of one or moreinflammatory mediators to their respective cognates. Alternatively, amodulator may alter, modify, reduce or eliminate the activity of one ormore inflammatory mediators by inhibiting the enzymatic activity, e.g.,phosphorylation activity, of the one or more inflammatory mediators.Alternatively, the one or more modulators may indirectly alter, modifyor eliminate the activity of one or more inflammatory mediators byattenuating the gene expression of one or more inflammatory mediators.In an aspect, the one or more modulators may indirectly alter oreliminate the activity of one or more inflammatory mediators byincreasing the expression of endogenous antagonists of the one or moreinflammatory mediators.

In an aspect, the one or more modulator can be a recombinant protein orpeptide or polynucleotide configured to modulate, alter, modify, orreduce the activity of the one or more inflammatory mediators. The oneor more modulator can be a polypeptide or nucleic acid molecule thateither induces expression of one or more anti-inflammatory mediators orattenuates expression of one or more pro-inflammatory mediators. The oneor more modulators can be a polypeptide or nucleic acid molecule thatagonizes or antagonizes binding of one or more inflammatory mediators toits cognate. In an aspect, the one or more modulators may be an antibodyor fragments thereof that block or modify the binding of an inflammatorymediator to its cognate. An example may be the chimeric monoclonalantibody infliximab (REMICADE® infliximab, Centocor Inc., Malvern, Pa.)that binds TNF-αand prevents it from binding to the TNF-α receptor.Alternatively, the one or more modulators may be all or part of one ormore soluble receptors that bind one or more inflammatory mediators andcompete for binding to the native receptors. An example may beetanercept, a soluble TNF-receptor (ENBREL® etanercept, Amgen, ThousandOaks, Calif.). The one or more modulator may be an anti-inflammatorymediator. For example, one or more anti-inflammatory mediators may bereleased into the peripheral blood of a subject to counterbalance theeffects of one or more pro-inflammatory inflammatory mediators. Anexample may be anakinra, a recombinant form of endogenous human IL-1receptor antagonist (IL-1Ra; KINERET® anakinra, Amgen, Thousand Oaks,Calif.).

Other examples of modulators of inflammatory mediators that are proteinsor peptides include, but are not limited to, adalimumab (anti-TNFantibody), abatacept (extracellular domain of cytotoxicT-lymphocyte-associated antigen 4, CTLA-4), alefacept (CD2 bindingportion of leukocyte-function-associated antigen-3 (LFA3) fused to humanIgG1), basiliximab and daclizumab (anti-IL-2 receptor alpha chainantibodies), efalizumab (anti-CD11a antibody), and recombinant forms ofinterferon-α, interferon-β, interferon-γ, and IL-2. In general, amodulator that is protein or peptide may be generated using standardrecombinant molecular biology techniques, for example, using thecorresponding cDNA sequences reported in GenBank as part of the NationalCenter for Biotechnology Information (NCBI). See, e.g., Benson, et al.,Nucleic Acids Res. 36:D25-D30, 2008, which is incorporated herein byreference. Antibodies to either an inflammatory mediator or itsrespective receptor may be generated using methods provided herein.

In an aspect, the reactive component can be a modulator that is a smallmolecule, an aptamer, or an inhibitory RNA, DNA, or nucleic acid.Modulators are contemplated that either directly or indirectly induce orattenuate expression of one or more inflammatory mediators and/oragonize or antagonize the activity of one or more inflammatorymediators. A small molecular inhibitor may inhibit a receptor or enzymethat is not itself an inflammatory mediator but is a component of thesignaling pathway that modulates expression of one or more inflammatorymediators. For example, the small molecule thalidomide (THALOMID®thalidomide, Celgene Corporation, Summit N.J.) inhibits TNF-α synthesiswhile modulating the levels of IL-10 and IL-12.

Other examples of small molecule modulators include, but are not limitedto, corticosteroids, e.g., hydrocortisone, cortisone, prednisone,prednisolone, methylprednisolone, meprednisone, triamcinolone,paramethsone, fluprednisolone, betametasone, and dexamethasone;nonsteroidal anti-inflammatory drugs (NSAIDS), e.g., selectivecycloxygenase (COX) inhibitors exemplified by celecoxib, etoricoxib,meloxicam, and valdecoxib and non-selective COX inhibitors exemplifiedby diclofenac, difluisal, etodolac, fenoprofen, fluripofen, ibuprofen,indomethacin, ketoprofen, ketorolac, nabumetone, naproxen, oxaprozin,piroxicam, sulindac, tenoxica, tiaprofen, tolmetin, azapropazone, andcarprofen; and disease modifying antirheumatic drugs (DMARDS), e.g.,methotrexate, azathioprine, pennicillamine, hydroxychloroquine,chloroquine, cyclophosphamide, cyclosporine, mycophenolate mofetil,gold, and sulfasalazine.

In an aspect, the one or more modulators of inflammatory mediators canbe a recombinant protein or peptide generated by one or more cellsincorporated into the device. The one or more cells can be geneticallymodified to synthesize and secrete the one or more modulators. Anextracorporeal device may deliver a therapeutic agent produced fromcells. See, e.g., U.S. Patent Publication 2007/0269489 A1, which isincorporated herein by reference. Cells that may be used for thispurpose include, but are not limited to, mammalian cells, enucleatedcells (e.g., erythrocytes), plants cells, bacteria, or yeast. DNAsequences corresponding to one or more modulators are cloned into anappropriate cell type using standard procedures with appropriateexpression vectors and transfection protocols. The genetically modifiedcells are encapsulated in one or more compartments of the bloodprocessing device and secrete the one or more modulators into theperipheral blood of a subject. The genetically modified cells are keptseparate from the circulation of a subject using a size-limitingbiocompatible mesh or membrane filter, for example, that allows passageof the one or more modulator, but not the larger cells.

In an aspect, endogenous cells associated with an inflammatory response,condition, disorder, or disease can be used to synthesize and secreteone or more modulators. Examples of endogenous cells associated with aninflammatory response, condition, disorder, or disease include, but arenot limited to, macrophages, dendrocytes, monocytes, T-lymphocytes,B-lymphocytes, neutrophils, eosinophils, basophils, and mast cells. Inan aspect, the one or more modulators are released into the peripheralblood of the subject to interact with endogenous immune cells currentlyin circulation and to trigger synthesis and/or secretion of one or moreother modulators from the circulating endogenous immune cells currently.In some aspects, the subject may be deficient in one or more types ofendogenous immune cells during the course of an inflammatory reaction.At least one endogenous cell type associated with the immune responsemay be encapsulated in one or more compartments of the device where thecells may secrete one or more modulators into the peripheral blood of asubject in response to appropriate activators.

In some aspects, the one or more modulators are released from syntheticvesicles or particles. Examples include any of a number of drug deliveryvehicles including, but not limited to, phospholipid vesicles(liposomes), nanoparticles, or hydrogels. The release of the one or moremodulators can be triggered by binding of a specific target to thesynthetic vesicle or particle. For example, one or more DNA aptamers maybe incorporated into hydrogel and designed to bind one or more specifictargets and release the contents of the hydrogel as described herein.

The device can include one or more reservoirs that store modulators ofone or more inflammatory mediators and release the modulators into thetreatment chamber and/or into the peripheral blood of the subject. Eachreservoir can contain one or more modulators. Release of the one or moremodulators from the one or more reservoirs into one or more treatmentchambers and/or into the peripheral blood is controlled by thecontroller component of the device. In some aspects, the one or moremodulators can be housed in multiple reservoirs associated with thedevice. For example, the device can include one or more microchips eachwith multiple reservoirs sealed with removable caps to enable controlledrelease of one or more inflammatory mediators. See, e.g., U.S. Pat. No.7,413,846; Maloney & Santini, Proceedings 26^(th) Annual InternationalConference IEEE EMBS, San Francisco, Calif., USA, Sep. 1-5, 2004, whichis incorporated herein by reference.

Energy sources can alter the functional structure of one or moreinflammatory mediators. The device can include one or more reactivecomponents that are one or more energy sources configured to alter thefunctional structure of one or more inflammatory mediators. The one ormore energy sources can be directed to peripheral blood within thetreatment chamber or can be directed outside the device to theperipheral blood. The one or more energy sources provide energy typesincluding, but not limited to, electromagnetic radiation, e.g.,ultraviolet, infrared, optical, microwave, or millimeter wave; acousticenergy, e.g., ultrasonic acoustic energy; heat; atmospheric pressureglow discharge; electron beam radiation; or gamma radiation. In someaspects, the energy source itself can alter the functional structure ofone or more inflammatory mediators. Alternatively, heat generated by theenergy source can alter the functional structure of one or moreinflammatory mediators.

The application of one or more energy sources to the peripheral blood inthe form of electromagnetic, acoustic, and or electronic energy caninduce denaturation and or degradation of one or more inflammatorymediators. An energy source can denature an inflammatory mediator byunfolding the structure and/or inducing changes in amino acid chainsand/or other side chains. The energy source can degrade an inflammatorymediator by cleaving one or more chemical bonds such as peptide bonds.The energy source can otherwise alter the functional structure of one ormore inflammatory mediators by inducing aggregation of the one or moreinflammatory mediators.

The device including the one or more energy sources can provide a set ofdiffering energy inputs specifically directed to altering the functionalstructure of one or more inflammatory mediators. The set of differingenergy inputs selectively resonates a plurality of resonant structuresin the one or more inflammatory mediators and can alter the functionalstructure of one or more inflammatory mediators. See, e.g., U.S. PatentApplication 2007/0021927 A1, which is incorporated herein by reference.The differing energy inputs are selected to resonate one or moreresonant structures among the group of proximate atoms comprising theone or more inflammatory mediators. Application of a series of differingenergy inputs can have a physical effect, such as transferringsubstantially more energy to a group of proximate atoms relative toother atoms in the surrounding medium, breaking a predetermined bondbetween two members of the group of proximate atoms, or changing akinetic parameter of a reaction involving a member of the group ofproximate atoms. The one or more resonant structures may be resonatedsimultaneously, sequentially, and/or in a temporally overlappingfashion. The series of differing energy inputs may be appliedsimultaneously, sequentially, and/or in a temporally overlappingfashion.

The set of differing energy inputs can be electromagnetic beams, each ofwhich can have one or more characteristics including, but not limitedto, a selected set of frequencies, a selected set of phases, a selectedset of amplitudes, a selected temporal profile, a selected set ofpolarizations, or a selected direction. The temporal profile of the setof differing energy inputs may be characterized by a selected beamduration, and/or by a selected change in frequency, modulationfrequency, phase, amplitude, polarization, or direction during aselected time interval. At least one electromagnetic beam may bepolarized, amplitude modulated, or frequency modulated, and it may be,for example, an infrared beam. A plurality of electromagnetic beams maydiffer in frequency, modulation frequency, phase, amplitude,polarization, or direction, and/or may intersect at a target location.The method may include scanning at least one electromagnetic beam.

In an aspect, the device can include reactive components that include anenergy field including, but not limited to, an electric field, amagnetic field, an electromagnetic field, a mechanical stress, amechanical strain, a lowered or elevated temperature, a lowered orelevated pressure, a phase change, an adsorbing surface, a catalyst, anenergy input, or a combination of any of these. The energy field canresult in degradation of the one or more inflammatory mediators.

The device including the one or more energy sources can generate heatthat induces denaturation of one or more inflammatory mediators.Exposure of most proteins or peptides to high temperature results inirreversible denaturation due to the weakening of long range bondsassociated with tertiary structure and cooperative hydrogen bondsassociated with helical structure. As these noncovalent bonds arebroken, the protein molecule becomes more flexible and exposed to thesolvent. Water molecules associated with the solvent form new hydrogenbonds that cannot be energetically overcome even as the protein moleculeis cooled, leaving the protein molecule in an altered or denaturedstate. The one or more inflammatory mediators may be inactivated bytreatment with a heat source. The heat source may be electrical, heatingthe entirety of the treatment chamber of the device. Alternatively, theheat source may heat a substrate of the device to which one or moreinflammatory mediators are attached. Alternatively, the heat source maybe more focused such as that experienced from exposure to focusedelectromagnetic energy, e.g. from a laser. In some aspects, focusedelectromagnetic energy may cause the substrate to bound to the one ormore inflammatory mediator to heat. In an aspect, the one or moreinflammatory mediators may bind to specific binding agents associatedwith one or more carbon nanotubes, the latter of which may emit heat inresponse to near infrared (NIR) radiation (Kam, et al., PNAS102:11600-11605, 2005, which is incorporated herein by reference.

The device including the one or more energy sources can generatemicrowave energy for use in altering the functional structure of one ormore inflammatory mediators. An energy source that incorporatesmicrowave energy can result in degradation of the one or moreinflammatory mediators. Microwaves are electromagnetic radiation withwavelengths between 0.01 and 1 meter and a frequency range between 0.3and 30 GHz. The efficiency of microwave denaturation and/or degradationcan be enhanced by including an acid, a protease, a chemical orcombination thereof. The effects of microwave energy on peptide bondintegrity can provide more that just rapid heating and suggests thatsome non-heat component of microwaves facilitates breakdown of thepeptide bond. See, e.g., Lill, et al., Mass. Spectrometry Rev.26:657-671, 2007, which is incorporated herein by reference.

The device including the one or more energy sources can generate focusedultrasound energy for use in altering the functional structure of one ormore inflammatory mediators. Sonication in the form of focusedultrasound energy may result in degradation of the one or moreinflammatory mediators. In an aspect, high intensity focused ultrasoundproduces cavitation bubbles that when collapsed yield very highlocalized pressures and high temperatures along with shear forces, jetsand shock waves. The local increase in temperature and pressure caneffectively denature proteins. See, e.g., Lopez-Ferrer, et al., J.Proteome Res. 7:3860-3867, 2008, which is incorporated herein byreference.

The device including the one or more energy sources can generate plasmaby atmospheric dielectric-barrier discharge for use in altering thefunctional structure of one or more inflammatory mediators. Plasmagenerated by atmospheric dielectric-barrier discharge may result indegradation of the one or more inflammatory mediators. See, e.g., Hou,et al., IEEE Transactions on Plasma Science 36:1633-1637, 2008, which isincorporated herein by reference. Plasma is an ionized gas in which acertain proportion of the electrons are free rather than bound to anatom or molecule. The plasma may generated by a non-thermal discharge atatmospheric pressure by application of high voltages across small gaps.The atmospheric dielectric-barrier discharge may be scalable from onemillimeter to one meter (Walsh, et al., IEEE Transactions on PlasmaScience 36:1314-1315, 2008, which is incorporated herein by reference. Aplasma jet and or a plasma brush may be used to degrade one or moreinflammatory mediators.

The device including the one or more energy sources can generate highenergy radiation for use in altering the functional structure of one ormore inflammatory mediators. High energy radiation may result indegradation of the one or more inflammatory mediators. Gamma radiationin the range of about 2.0 kGy to about 23.0 kGy is able to denatureprotein in a dose dependent manner as determined by size chromatography.See, e.g., Vuckovic, et al., J. Serb. Chem. Soc. 70:1255-1262, 2005,which is incorporated herein by reference. Sources of gamma radiationthat may be included in the device include but are not limited tocobalt-60, cesium-137, and technetium-99.

The device including the one or more energy sources can generateelectron beam radiation for use in altering the functional structure ofone or more inflammatory mediators. Electron beam radiation can resultin degradation of the one or more inflammatory mediators. Electron beamenergy of 92.9 kGy induces physical changes and loss of proteinantigenicity (Katial, et al., J. Allergy Clin. Immunol. 110:215-219,2002, which is incorporated herein by reference. A nanoscale electronbeam generator may be devised from a network array structure of carbonnanotubes. See, e.g., U.S. Pat. No. 7,355,334, which is incorporatedherein by reference.

Two or more reactive components can be combined to alter the functionalstructure of one or more inflammatory mediators. The device can includetwo or more reactive components that have been combined to alter thefunctional structure of one or more inflammatory mediators. The two ormore combined reactive components can be one or more binding agentcombined with one or more denaturing agent, degradative agent,modulator, energy source, or combination thereof. For example, a bindingagent, e.g., oligonucleotide aptamer, may be used to capture one or moreinflammatory mediators in the treatment chamber of the device prior totreatment with a denaturing agent, degradative agent, modulator, energysource, or a combination thereof.

In an aspect, the two or more reactive components of the device can beincorporated into a single biomolecule. For example, the first reactivecomponent can be a binding agent, e.g., an antibody, that includes asecond reactive component that is a degradative activity. Certainantibodies are capable of cleaving the amide bond of peptide bonds. See,e.g., Janda, et al., Science 241:1188-1191, 1988; Lacroix-Desmazes, etal., J. Immunol. 177:1355-1365, 2006; Ponomarenko, et al., PNAS,103:281-286, 2006; and U.S. Pat. No. 6,387,674, which are incorporatedherein by reference. Alternatively, the first reactive component can bea binding agent, e.g., an antibody, and includes a second reactivecomponent that is a reactive oxygen species. The one or moreinflammatory mediators can bind to one or more binding agents that arecatalytic antibodies capable of generating the reactive oxygen speciesH₂O₂ in response to UV radiation. See, e.g., Wentworth, et al., Science293:1806-181811, 2001; Wentworth, Science 296:2247-2248, 2002;Wentworth, et al., PNAS, 97:10930-10935, 2000, which are incorporatedherein by reference. One or more antibodies or other binding agents canbe generated for both binding and degradation of one or moreinflammatory mediators.

In another aspect, the two or more reactive components of the device canbe incorporated into a single biomolecule and can include a firstcomponent that is a binding agent, e.g., an aptamer, and a secondcomponent that is a degradative agent, e.g., a protease. For example,one or more proteases may be conjugated or chemically linked to one ormore oligonucleotide-based aptamers. The oligonucleotide-based aptamersare designed to bind one or more inflammatory mediators. Upon binding tothe oligonucleotide-based aptamers, the one or more inflammatorymediators are brought into proximity to the one or more proteasesresulting in proteolytic degradation of the one or more inflammatorymediators. Examples of proteases have been provided herein and may belinked to oligonucleotide-based aptamers using any of a number ofmethods for conjugating a polypeptide to an oligonucleotide. In afurther aspect, a polypeptide protease may be conjugated to anoligonucleotide-based aptamer using a streptavidin-biotin bridge byintroducing a biotinylated oligonucleotide into the aptamer sequence andlinking it to a biotinylated protease through a streptavidin bridge.Alternatively, the polypeptide protease may be conjugated to theoligonucleotide-based aptamer using a thiol-maleimide linkage in which acarbon with an attached thiol group is placed on the aptamer and reactswith a maleimide group added to the C terminus of the protease. See,e.g., Nitin, et al., Nucleic Acids Res. 32:e58, 2004, which isincorporated herein by reference. A number of modified nucleotides arecommercially available for use in synthesizing oligonucleotide aptamerswith amines or other side chains for cross-linking (TriLinkBiotechnologies, San Diego, Calif.; Sigma Aldrich, St. Louis, Mo.).

In a further aspect, the first reactive component can be a binding agentlinked to a second reactive component encapsulated in a tunable vesicle.For example, the second reactive component, e.g., a denaturing and/ordegradative agent, can be encapsulated in a tunable hydrogel. Thebinding of one or more inflammatory mediators to the first reactivecomponent, e.g., binding agent, releases the denaturing and/ordegradative agent from the hydrogel. In an aspect, target-responsivehydrogels may be generated in which the contents of the hydrogel areselectively released in response to binding a specific target. Thehydrogel may incorporate one or more binding agents that are antibodies.The hydrogel may release its contents in response to an antibody-antigeninteraction. See, e.g., Miyata, et al., PNAS 103:1190-1193, 2006, whichis incorporated herein by reference. In an aspect, the target-responsivehydrogel may incorporate one or more binding agents that areoligonucleotide-based aptamers and release its contents in response toan aptamer-ligand interaction. See Yang, et al., J. Am. Chem. Soc.130:6320-6321, 2008, which is incorporated herein by reference. In thelatter case, two or more distinct aptamers configured to partiallyoverlap during hybridization may be copolymerized into a polyacrylamidehydrogel. At least one of the two or more aptamers further binds to aspecific target, e.g., an inflammatory mediator. When the inflammatorymediator binds to the aptamer, the number of nucleotide bases availablefor hybridization between the overlapping aptamers is reduced, causingthem to separate. This separation allows the hydrogel to dissolute andrelease its contents. A target responsive hydrogel may be generatedwhich incorporates aptamers that specifically recognize one or moreinflammatory mediators. The hydrogel itself may be loaded with one ormore proteases or other reactive components that are configured to alterthe functional structure of inflammatory mediators. The contents of thehydrogel are released upon binding of the one or more inflammatorymediators to the aptamers associated with the hydrogel. In a furtheraspect, hydrogels may be used for molecular imprinting. See, e.g., Byrneet al., “Molecular imprinting within hydrogels,” Advanced Drug DeliveryReviews, 54: 149-161, 2002, which is incorporated herein by reference.

Substrates for Reactive Components

The one or more reactive components including binding agents, denaturingagents, degradative agents, modulators, or combinations there of can befree in solution within one or more treatment chambers of the device.Alternatively, the one or more reactive components can be immobilized ona solid substrate within the one or more treatment chambers of thedevice. The solid substrate may be a matrix, e.g., a bead or filter,that is added to one or more treatment chambers of the device. Examplesof applicable solid substrates include, but are not limited to, beads,particles, membranes, semi-permeable membranes, capillary, ormicroarrays. The solid substrate may be comprised of an inorganicmaterial, e.g., glass, alumina, silica, silicon, zirconia, graphite,magnetite, semiconductors, or combinations thereof. Alternatively, thesolid substrate may be comprised of an organic material, e.g.,polysaccharides including agarose, dextran, cellulose, chitosan, andpolyacrylamide, polyacrylate, polystyrene, polyvinyl alcohol, orcombinations thereof. Alternatively, the one or more specific bindingagents or one or more reactive components may be associated with a solidsubstrate that are cells, e.g., mammalian cells, enucleatederythrocytes, bacteria, or viral particles or vesicles such as liposomesor other micellular vesicles.

In some aspects, the one or more reactive components, either free insolution or bound to a solid substrate, can be prevented from leavingthe one or more treatment chambers of the device either due to sizeexclusion using a filter or mesh or due to physical attachment to thedevice. In a detailed aspect, one or more inflammatory mediators presentin the blood can bind to the one or more reactive components as theblood passes through the device and sequestered for inactivation.Alternatively, the one or more reactive components, either free insolution or bound to a solid substrate, can be released into the bloodstream and allowed to bind one or more inflammatory mediators while incirculation. In this aspect, the one or more reactive components can berecaptured by the device and the functional structure of the one or morebound inflammatory mediators can be altered.

The one or more reactive components can be bound to the solid substrateeither directly or indirectly. For example, the one or more reactivecomponents may be coupled to the solid substrate by covalent chemicalbonds between particular functional groups on the specific binding agent(e.g., primary amines, sulfhydryls, carboxylic acids, hydroxyls, andaldehydes) and reactive groups on the solid substrate. A variety ofactivating compounds and schemes for directly bonding ligands to solidsubstrates are known. Some examples include, but are not limited to,cyanogen bromide, cyanuric chlorde, epichlorohydrin, divinyl sulphone,p-toluenesulphonyl chloride, 1,1′-carbonyldiimidazole, sodiummeta-periodate, 2-fluoro-1-methylpyridiniumtoluene-4-sulphonate,glycidoxypropyl-trimethoxysilane and 2,2,2-trifluoroethanesulphonylchloride. For example, cyanogen bromide in base reacts with hydroxyl(OH) groups on agarose solid substrate to form cyanate esters orimidocarbonates. These groups readily react with primary amines undermild conditions resulting in a covalent coupling of the ligand to theagarose solid substrate. Reactive imidocarbonates may also be formed oncarbon nanotubes, for example, through reactive carboxyl groupsgenerated by treatment of the nanotubes with oxidizing agents. See,e.g., Bianco, et al., in Nanomaterials for Medical Diagnosis andTherapy. pp. 85-142. Nanotechnologies for the Live Sciences Vol. 10Edited by Challa S. S. R. Kumar, WILEY-VCH Verlag GmbH & Co. KGaA,Weinheim, 2007, which is incorporated herein by reference.Functionalization of silicon chips with carboxyl groups can besubsequently used to immobilize proteins in the presence ofN-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) andN-hydroxysuccinimide ester (NHS). See, e.g., Hu, et al., Rapid Commun.Mass Spectrom. 21:1277-1281, 2007, which is incorporated herein byreference.

The one or more reactive components may or may not have linking orspacer groups bound to the C-terminus which when present may be used tobind the specific binding agent to the solid substrate indirectly. Whenpresent the linking group may be a polymer or a monomer. A linking groupmay be a chain of from 1-10 amino acids. Other examples of linkinggroups include, but are not limited to, polyethylene glycol,polypropylene glycol, polyesters, polypeptides, polyethers,polysaccharides, glycidoxyalkyl, alkoxyalkyl, alkyl, glycidoxypropyl,ethyl, propyl, phenyl and methacryl; and silicon containing linkinggroups such as diethyl(triethoxysilylpropyl)malonate;3-mercaptopropyltrimethoxysilane; 3-aminopropyltrimethoxysilane;N-[(3-trimethoxysilyl)propyl]ethylenediaminetriacetic acid;p-(chloromethyl)phenyltrimethoxysilane; vinyltriethoxysilane;3-bromopropyltriethoxysilane; and 3-glycidoxypropyltrimethoxysilane.

In general, any of a number of homobifunctional, heterofunctional,and/or photoreactive cross linking agents may be used to conjugate oneor more reactive components to an appropriately derivatized substrate.Examples of homobifunctional cross linkers include, but are not limitedto, primary amine/primary amine linkers such as BSOCES((bis(2-[succinimidooxy-carbonyloxy]ethyl)sulfone), DMS (dimethylsuberimidate), DMP (dimethyl pimelimidate), DMA (dimethyl adipimidate),DSS (disuccinimidyl suberate), DST (disuccinimidyl tartate), Sulfo DST(sulfodisuccinimidyl tartate), DSP (dithiobis(succinimidyl propionate),DTSSP (3,3′-dithiobis(succinimidyl propionate), EGS (ethylene glycolbis(succinimidyl succinate)) and sulfhydryl/sulfhydryl linkers such asDPDPB (1,4-di-(3′-[2′pyridyldithio]-propionamido) butane). Examples ofheterofunctional cross linkers include, but are not limited to, primaryamine/sulfhydryl linkers such as MBS(m-maleimidobenzoyl-N-hydroxysuccinimide ester), Sulfo MBS(m-maleimidobenzoyl-N-hydroxysulfosuccinimide), GMBS(N-gamma-maleimidobutyryl-oxysuccinimide ester), Sulfo GMBS(N-γ-maleimidobutyryloxysulfosuccinimide ester),EMCS(N-(epsilon-maleimidocaproyloxy) succinimide ester), SulfoEMCS(N-(epsilon-maleimidocaproyloxy) sulfo succinimide), SIAB(N-succinimidyl(4-iodoacetyl)aminobenzoate), SMCC (succinimidyl4-(N-maleimidomethyl) cyclohexane-1-carboxylate), SMPB (succinimidyl4-(rho-maleimidophenyl) butyrate), Sulfo SIAB(N-sulfosuccinimidyl(4-iodoacetyl)aminobenzoate), Sulfo SMCC(sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate),Sulfo SMPB (sulfosuccinimidyl 4-(rho-maleimidophenyl) butyrate), andMAL-PEG-NHS (maleimide PEG N-hydroxysuccinimide ester);sulfhydryl/hydroxyl linkers such as PMPI (N-rho-maleimidophenyl)isocyanate; sulfhydryl/carbohydrate linkers such as EMCH(N-(epsilon-maleimidocaproic acid) hydrazide); and amine/carboxyllinkers such as EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimidehydrochloride).

In some aspects, the one or more reactive components can be linked to asolid substrate through non-covalent interactions. Examples ofnon-covalent interactions include, but are not limited to,protein-protein interactions such as those between avidin/streptavidinand biotin, protein A and immunoglobulins, protein G andimmunoglobulins, or secondary antibodies with primary antibodies. Forexample, the one or more reactive components may be modified with biotinusing standard methods and bound to a solid substrate derivatized withstreptavidin. Alternatively, one or more reactive components may bemodified with streptavidin and bound to a solid substrate derivatizedwith biotin. A single chain antibody may incorporate streptavidin aspart of a fusion protein. See, e.g., Koo, et al. Appl. Environ.Microbiol. 64:2497-2502, 1998) to facilitate attachment of the antibodyto the solid substrate via a biotin-streptavidin linkage. Solidsubstrates such as beads or other particulate substrates derivatizedwith protein A, protein G, streptavidin, avidin, biotin, secondaryantibodies are available from commercial sources (from, e.g.,Pierce-Thermo Scientific, Rockford, Ill., Sigma-Aldrich, St. Louis,Mo.). In some aspects, the one or more reactive components may bind tothe solid substrate through a non-covalent interaction and be furthercross-linked to the solid substrate using a cross-linking agent.

In an aspect, the one or more reactive components can be associated witha solid substrate that are cells, e.g., mammalian cells, enucleatederythrocytes, bacteria, or viral particles, or vesicles such asliposomes or other micellular vesicles. Cells and vesicles may bemodified with one or more reactive components using many of the samemethods as provided herein. One or more reactive components may be boundto cells or vesicles using one or more homobifunctional orheterofunctional cross-linkers through primary amines and carboxylgroups. Alternatively, cells may be modified with one or more reactivecomponents using a biotin-streptavidin bridge. For example, one or morereactive components may be biotinylated and linked to a non-specificallybiotinylated cell surface through a streptavidin bridge. An antibody,aptamer, or receptor may be biotinylated using standard procedures. Thesurface membrane proteins of a cell may be biotinylated using an aminereactive biotinylation reagent such as, for example, EZ-LinkSulfo-NHS-SS-Biotin (sulfosuccinimidyl2-(biotinamido)-ethyl-1,3-dithiopropionate; Pierce-Thermo Scientific,Rockford, Ill., USA; see, e.g., Jaiswal, et al. Nature Biotech.21:47-51, 2003; U.S. Pat. No. 6,946,127).

In an aspect, the one or more reactive components can be associated withlipid or micellular vesicles. In some aspects, the one or more reactivecomponents can be antibodies attached to a liposome. Antibodies may beadded to liposomes using cross-linking agents and protein A. See, e.g.,Renneisen, et al., J. Bio. Chem., 265:16337-16342, 1990, which isincorporated herein by reference. The liposomes are formed from drylipid in the presence of an aqueous solution, e.g., a buffer ofappropriate pH followed by extrusion through a high pressure devicefitted with a polycarbonate filter with the desired pore size to formliposomes of a specific size range. The liposomes are modified withN-succinimidyl 3-(2-pyridyldithio) propionate-modified protein A. Theone or more antibodies are linked to the liposomes through selectivebinding to the protein A. Alternatively, thiolated antibodies may becovalently linked to liposomes prepared with 4-(p-maleimidophenyl)butyrylphosphatidyl-ethanolamine. See, e.g., Heath, et al., PNAS80:1377-1381, 1983, which is incorporated herein by reference.

In an aspect, the one or more reactive components can be one or moremultispecific antibody complex with one or more components thatrecognize an antigen on the surface of a cell or vesicle and a secondone or more components that recognize one or more inflammatorymediators. For example, a multispecific antibody complex may have onecomponent that binds to erythrocytes and a second component that bindsone or more inflammatory mediators. A number of examples ofmultispecific antibodies that recognize erythrocytes and variousendogenous targets or pathogens are provided. See, e.g., U.S. Pat. Nos.5,470,570 and 5,843,440; U.S. Pat. App. Nos. 2003/0215454 A1 and2006/0018912 A1.

In some aspects, the one or more reactive components can be expressed onthe surface of a cell. The one or more reactive components may benaturally expressed on the surface of a cell, such as a receptor of aspecific inflammatory mediator on a specific cell type. Alternatively,the one or more reactive components may be expressed on the surface of acell using genetic manipulation. For example, cells may be geneticallymanipulated to express a receptor that binds one or more inflammatorymediators. In one example, the genetic expression of a receptor on thesurface of Chinese hamster ovary (CHO) cells is capable of binding thecytokine interleukin 1 (IL-1). See, e.g., Curtis, et al., PNAS USA86:3045-3049, 1989, which is incorporated herein by reference.Alternatively, cells may be genetically manipulated to express one ormore specific antibodies on the cell surface. Methods have been providedfor cell surface expression of single chain Fv antibody fragments (scFv)fused to membrane-associated proteins. See, e.g., Ho, et al., Proc.Natl. Acad. Sci. USA 103:9637-9642, 2006; Francisco, et al., Proc. Natl.Acad. Sci. USA 90:10444-10448, 1993; U.S. Pat. Appl. No. 2006/0083716,which are incorporated herein by reference. In this aspect, the cDNAsequence encoding all or part of an inflammatory mediator-specificantibody is fused in an expression construct in frame with amembrane-associated protein and expressed in an appropriate cell type.

EXEMPLARY ASPECTS Example 1

Intracorporeal Device for Sensing, Binding and Altering InflammatoryMediators

A method for treating an inflammatory condition or disease is providedand includes an intracorporeal device designed to sense one or moreinflammatory mediators in peripheral blood of a subject and a modulatingmeans configured to bind the one or more inflammatory mediators andalter the functional structure or activity of the one or moreinflammatory mediators to achieve a desired target value. Theintracorporeal device includes a controller in communication with thesensor and configured to adjust the modulating means to achieve thetarget value of the detected one or more inflammatory mediators in theperipheral blood of the subject. The device optionally includes areceiver for receiving and processing data regarding the sensed levelsof one or more inflammatory mediators and a transmitter for transmittingdata regarding the sensed levels of one or more inflammatory mediatorsto an external controller.

The intracorporeal device is placed in or proximal to one or more bloodvessels of a subject. In this example, the intracorporeal device is ahollow stent-like structure that is placed into a vessel at or near thesite of inflammation using a catheter guide wire. The components of theintracorporeal device including sensors, controller, binding elements,and reactive components are affixed to and/or incorporated into one orboth surfaces of the stent-like structure. The intracorporeal device isconfigured such that blood in the vessel is allowed to flow through itessentially unobstructed.

The intracorporeal device includes one or more sensors that sense thelevels of one or more inflammatory mediators in the peripheral blood ofa subject. In this example, the one or more sensors are piezoelectricsensors in which aptamers directed against the proinflammatory mediatorsTNF-α and IL-1 are used as recognition elements. The interaction ofTNF-α and IL-1 with their respective recognition elements triggers thepiezoelectric sensor to send a signal to the controller. In thisexample, the controller is an integral component of the intracorporealdevice. The controller calculates the levels of TNF-α and/or IL-1 in theperipheral blood based on the input from the sensors and compares thesedata with target values, e.g., desired concentrations of TNF-α and/orIL-1. In some instances, the target value for TNF-α and/or IL-1 is thatobserved in a normal subject not experiencing an inflammatory disease ora disease resulting in an inflammatory response. In other instances, thetarget value for TNF-α and/or IL-1 may represent a reduction of at least20%, at least 40%, at least 60%, at least 80%, or at least 100% relativeto the current level of TNF-α and/or IL-1 in the peripheral blood of thesubject. The controller may optionally send a wireless signal to anexternal controller to alert the subject and/or one or more caregiversas to the levels of TNF-α and/or IL-1 in the peripheral blood of thesubject.

The intracorporeal device further includes one or more binding agentsfor capturing one or more inflammatory mediators within the treatmentchamber of the intracorporeal device. In this example, the bindingagents are antibodies directed against the proinflammatory mediatorsTNF-α and IL-1. Antibodies to TNF-α, IL-1, and many other inflammatorymediators are available from commercial sources (from, e.g., NovusBiological, Littleton, Colo.; Sigma-Aldrich, St. Louis, Mo.; UnitedStates Biological, Swampscott, Mass.) or are readily generated usingstandard methods. The antibodies directed against TNF-α and IL-1 arepreferably incorporated within the stent-like structure of theintracorporeal device at one or more sites to maximize exposure to TNF-αand IL-1 in the blood of a subject. Protocols are provided forchemically linking an antibody to a collagen-coated stent usingN-succinimidyl-3-(2-pyridyldithiol)-propionate as a cross-linker. See,e.g., Jin, et al., J. Gene Med. 8:786-793, 2008, which is incorporatedherein by reference.

The intracorporeal device further includes one or more reactivecomponents designed to alter the functional structure of one or moreinflammatory mediators and as such modulate their physiological effectsto achieve the desired target values. In some aspects, the reactivecomponent can be the one or more specific binding agents capable ofdirectly altering the functional structure of the one or moreinflammatory mediators. In this example, the binding agent antibodiesdirected against TNF-α and IL-1 have intrinsic catalytic activitytriggered by a controllable energy source. In response to ultravioletenergy released by the intracorporeal device within the treatmentchamber, the TNF-α and IL-1 binding agent antibodies produce reactiveoxygen species that alter the functional structure of TNF-α and IL-1.The release of ultraviolet energy in the treatment chamber is triggeredby the controller based on the sensed levels of TNF-α and IL-1 in theperipheral blood. In the absence of a triggering event, the TNF-α andIL-1 bound to the binding agent antibodies will eventually dissociate inan intact and active form and return to the peripheral blood of thesubject.

Example 2

Intracorporeal Device for Sensing and Altering Inflammatory Mediators

A method for treating an inflammatory condition or disease is providedand includes an intracorporeal device designed to sense one or moreinflammatory mediators in the peripheral blood of a subject and amodulating means to alter the activity of the one or more inflammatorymediators to achieve a desired target value. The intracorporeal deviceis placed in or proximal to one or more blood vessels and includes atreatment chamber configured to receive at least a portion of theperipheral blood through a flow route, the controller configured tocontrol flow of peripheral blood through the flow route into thetreatment chamber. The treatment chamber includes one or more reactivecomponents utilizing a set of differing energy inputs specificallydirected to the one or more inflammatory mediators within a treatmentchamber of the device and configured to alter a functional structure ofthe one or more inflammatory mediators in the peripheral blood. Thedevice optionally includes a transmitter configured to transmit thesensed levels of one or more inflammatory mediators to an externalcontroller.

The intracorporeal device includes one or more sensors that sense thelevels of one or more inflammatory mediators in the peripheral blood ofa subject. In this example, the one or more sensors are aptamer-basedmolecular beacons designed to fluoresce in response to selectivelybinding one or more inflammatory mediators. The aptamer-based molecularbeacons include a recognition element and at least one fluorescingmoiety and at least one quenching moiety. The recognition elementselectively interacts with one or more proinflammatory mediators, e.g.,TNF-α and IL-1 and one or more anti-inflammatory mediators, e.g., IL-10.Fluorescence induced by electromagnetic energy emitted by theintracorporeal device is quenched in the absence of TNF-α, IL-1, orIL-10. The binding of TNF-α, IL-1, and/or IL-10 to the respectiveselective aptamer induces a conformational change in the aptamer andincreases the distance between the fluorescing moiety and the quenchingmoiety resulting in a fluorescent signal in response to electromagneticenergy. The level of fluorescent signal is proportional to the level ofTNF-α, IL-1, and/or IL-10 in the blood sample. The emitted fluorescenceis captured by a CCD or CMOS detector and a corresponding signal is sentto the controller.

The controller calculates the levels of TNF-α, IL-1, and/or IL-10 in theperipheral blood based on the input from the sensors and compares thesedata with target values, e.g., desired concentrations of TNF-α, IL-1,and/or IL-10. In some instances, the target value can represent a ratioof concentrations for the proinflammatory mediators TNF-α and IL-1relative to the anti-inflammatory mediator IL-10. The controller mayoptionally send a wireless signal to an external controller to alert thesubject and/or one or more caregivers as to the levels of TNF-α, IL-1,and/or IL-10 in the peripheral blood of the subject.

The controller optionally controls flow of the peripheral blood into thetreatment chamber. The controller initiates release of one or more setsof differing energy inputs into the treatment chamber of theintracorporeal device to specifically alter a functional structure ofone or more inflammatory mediators, e.g., TNF-α, IL-1, and/or IL-10. Inthis example, the set of differing energy inputs selectively resonates aplurality of resonant structures in TNF-α, IL-1, and/or IL-10. Theresonance can alter the functional structure of TNF-α, IL-1, and/orIL-10 by transferring substantially more energy to at least a portion ofthe group of proximate atoms than to other atoms in the medium, breakinga predetermined bond between two members of the group of proximateatoms, and/or changing a kinetic parameter of a reaction involving amember of the group of proximate atoms. The sets of differing energyinputs are transmitted in a simultaneous, sequential, and/or in atemporally overlapping pattern that specifically disrupts oneinflammatory mediator relative to another in the treatment chamber. Theenergy inputs are electromagnetic energy emitted at one or morewavelengths from miniaturized diode lasers associated with theintracorporeal device. Treatment continues until the target values forthe one or more inflammatory mediators have been achieved.

Example 3

Intracorporeal Device for Sensing Inflammatory Mediators andControllably Releasing Inflammatory Modulators

A method for treating an inflammatory condition or disease is providedincluding an intracorporeal device designed to sense one or moreinflammatory mediators in the peripheral blood of a subject and areservoir to controllably release one or more reactive components, e.g.,one or more inflammatory modulators, in the peripheral blood. The one ormore inflammatory modulators are configured to alter a functionalstructure of the one or more inflammatory mediators to achieve a desiredtarget value of the one or more inflammatory mediators in the peripheralblood. The intracorporeal device is placed in or proximal to one or moreblood vessels of a subject and includes one or more sensors for sensingone or more inflammatory mediators, a controller to receive dataregarding the sensed levels of one or more inflammatory mediators, oneor more reservoirs containing one or more inflammatory modulatorscontrollably opened or closed by the controller. The device mayoptionally a transmitter configured to transmit the sensed levels of theone or more inflammatory mediators to an external controller.

The intracorporeal device can be a stent-like structure that resides ina fixed position in a blood vessel. Alternatively, the intracorporealdevice can be free to travel in the lumen of one or more blood vessels.See, e.g., U.S. Patent Application 2007/0156211 A1, which isincorporated herein by reference.

The intracorporeal device includes one or more sensors for sensing thelevels of one or more inflammatory mediators in the peripheral blood ofa subject. The sensors include one or more recognition elements designedto sense levels of one or more inflammatory mediators. In this example,the recognition elements associated with the sensors are antibodiesdirected towards the proinflammatory mediators TNF-α and IL-1 and theanti-inflammatory mediator IL-10. As blood passes by the intracorporealdevice, the sensors associated with the device respond to the presenceof TNF-α, IL-1 and/or IL-10 by sending an electrical signal to acontroller. In this example, the controller is an integral component ofthe intracorporeal device. The controller compares the data regardingthe sensed level of TNF-α, IL-1 and/or IL-10 in the peripheral blood ofa subject with target values, e.g., a desired range of concentrations ofthe proinflammatory or anti-inflammatory mediators. In some instances,the target value can represent a ratio of concentrations, for example,the concentration of proinflammatory mediators TNF-α and IL-1 relativeto the concentration of the anti-inflammatory mediator IL-10. Asappropriate, the controller signals release of one or more inflammatorymodulators from reservoirs associated with the intracorporeal device toachieve the target values.

The intracorporeal device includes one or more micro-reservoirscontaining the one or more inflammatory modulators. The micro-reservoirsmay be covered by a seal that is disrupted in response to an electricalsignal from the controller. See, e.g., U.S. Pat. No. 7,413,846; Maloney& Santini, Proceedings 26^(th) Annual International Conference IEEEEMBS, San Francisco, Calif., USA, Sep. 1-5, 2004, which are incorporatedherein by reference. The inflammatory modulators in the micro-reservoirsmay directly or indirectly modulate the level of one or moreinflammatory mediators. In this example, the inflammatory modulatorsdirectly alter the functional levels of TNF-α, IL-1 and/or IL-10 and arecontained in separate reservoirs within the intracorporeal device. Oneset of reservoirs contains a soluble TNF-α receptor, e.g., etanercept. Asecond set of reservoirs contains an IL-1 receptor antagonist, e.g.,anakinra. The third set of reservoirs contains recombinant IL-10.Release of the inflammatory modulators from the one or more reservoirsis controlled by signals sent from the controller and is based on thesensed level of TNF-α, IL-1 and/or IL-10 in the peripheral blood. Inthis configuration, the levels of the proinflammatory mediators TNF-αand IL-1 may be controllably decreased and the levels of theanti-inflammatory mediator IL-10 may be controllably increased toachieve appropriate target values of each inflammatory mediator.

Example 4

Intracorporeal Device for Sensing Inflammatory Mediators andAutomatically Releasing Inflammatory Modulators

A method for treating an inflammatory condition or disease is providedand includes an intracorporeal device designed to sense one or moreinflammatory mediators in the peripheral blood of a subject and toautomatically release one or more reactive components, e.g., one or moreinflammatory modulators, in response to the sensed condition to achievea desired target value of the one or more inflammatory mediators. Theintracorporeal device is placed in or proximal to one or more bloodvessels of a subject and includes one or more sensors for sensing one ormore inflammatory mediators, one or more reservoirs directly linked tothe sensors for immediate and automatic release of one or moreinflammatory modulators, and optionally a transmitter for transmittingthe sensed levels of one or more inflammatory mediators to an externalcontroller. Data regarding the level of one or more inflammatorymediators in the peripheral blood of a subject is optionally transmittedto an external controller for monitoring by the subject and/or one ormore caregivers.

The intracorporeal device includes one or more sensors directly linkedto release of one or more inflammatory modulators in response to sensingone or more inflammatory mediators. For example, the one or more sensorsmay be associated with target responsive vesicles that release thevesicle contents in response to binding one or more inflammatorymediators. In this example, the one or more sensors include recognitionelements that interact with proinflammatory mediators, e.g., TNF-α,IL-1, IL-6, and IL-8. The recognition elements are aptamers that bindTNF-α, IL-1, IL-6, and/or IL-8 and are also involved in encapsulatingone or more inflammatory modulators into target responsive vesicles. Forexample, two distinct aptamers capable of partial overlappinghybridization are copolymerized into a polyacrylamide-based hydrogel.One of the two aptamers is a recognition element that binds to aproinflammatory mediator, e.g., IL-1. The interaction of IL-1 with theaptamer recognition element causes the two partially overlappingaptamers to separate from one another and to change the properties ofthe hydrogel, resulting in release of the contents of the hydrogel. Thehydrogel is loaded with one or more inflammatory modulator, e.g., anIL-1 receptor antagonist. In this aspect, an increase in IL-1 in theperipheral blood of a subject leads to increased binding to theaptamer/hydrogel complex, separation of the aptamer pair, and increasedrelease of the IL-1 receptor antagonist. The target-responsive vesiclesare retained in one or more compartments of the intracorporeal device.The compartments include a membrane or screen through which inflammatorymediators and inflammatory modulators may diffuse but otherwise preventsthe release of the target-responsive vesicles into the peripheralcirculation.

The interaction of one or more inflammatory mediators with the targetresponsive vesicles can be monitored using fluorescence resonance energytransfer (FRET). For example, the two overlapping aptamers incorporatedinto the hydrogel can be modified with a fluorescence emitting molecule,e.g., AF 647 (Molecular Probes-Invitrogen, Carlsbad, Calif.) and afluorescence quenching molecule, e.g., QSY 21 (MolecularProbes-Invitrogen, Carlsbad, Calif.). In the absence of binding aproinflammatory mediator, e.g., IL-1, the two aptamers remain linked toone another, fluorescence associated with the aptamers remains quenchedand the inflammatory modulator remains encapsulated in the hydrogel.Upon binding IL-1, the aptamers change configuration relative to oneanother separating the fluorescence emitting molecule and the quenchingmolecule resulting in a measurable fluorescent signal. The fluorescentsignal is measured by a CCD device or other light capture device withinthe intracorporeal device and transmitted wirelessly to an externalcontroller. Alternatively, the fluorescent signal is measured throughthe skin using near infrared imaging and fluorescent dyes or quantumdots that fluoresce in the near infrared wavelengths. See, e.g.,Frangioni Curr. Opin. Chem. Biol. 7:626-634, 2003, which is incorporatedherein by reference. An example of a near infrared dye FRET pair wouldbe IRDye 800CW and IRDye QC-1 Quencher (LI-COR, Lincoln, Nebr.).

Example 5

Extracorporeal Device for Sensing and Sequestering InflammatoryMediators

A method for treating an inflammatory disease is provided and includesan extracorporeal device configured to sense and sequester one or moreinflammatory mediators in the peripheral blood of a subject to achieve adesired target value. The extracorporeal device includes a flow routefor extracting blood from a subject, a component for optionallyfractionating the whole blood into separate components, one or moresensors for sensing the levels of one or more inflammatory mediators, acontroller to receive data regarding the sensed levels of one or moreinflammatory mediators, one or more binding elements incorporated intoone or more treatment chambers to sequester one or more inflammatorymediators, and a flow route for returning the processed blood to thesubject.

Blood is withdrawn from a subject through a catheter inserted into avein or artery of the subject. The catheter is further attached to oneor more conduits that flow into the extracorporeal device. The wholeblood is fractionated by centrifugation and/or filtration within theextracorporeal blood processing device to isolate the plasma.Alternatively, whole blood from a subject is fractionated usingcommercially available apheresis instrumentation (from, e.g.,CardianBCT, CO, USA; Baxter, Ill., USA) and the resulting plasma is sentfrom the apheresis instrument to the extracorporeal device for furtherprocessing.

The whole blood or plasma is passed over one or more sensors associatedwith the extracorporeal device. The one or more sensors are configuredto sense one or more inflammatory mediators and include one or morerecognition elements. In this example, the recognition elements areantibodies configured to selectively recognize the proinflammatorymediators, e.g., TNF-α, IL-1, IL-6, and IL-8. The recognition elementantibodies are immobilized on one or more solid substrates, e.g., a CMSsensor chip (Biacore, Inc.—GE Healthcare, Piscataway, N.J.), bycrosslinking free amino acid groups associated with the antibodies toN-hydroxylsuccinimide carbodiimide-activated carboxyl groups associatedwith the sensor chip. The levels of TNF-α, IL-1, IL-6, and/or IL-8 inthe whole blood or plasma are sensed via surface plasmon resonance. Anexample of instrumentation that uses surface plasmon resonance is theBIACORE system which includes a sensor microchip, a laser light sourceemitting polarized light, an automated fluid handling system, and adiode array position sensitive detector. See, e.g., Biacore, Inc.—GEHealthcare, Piscataway, N.J.; and Raghavan & Bjorkman Structure3:331-333, 1995, which are incorporated herein by reference.

Data regarding the levels of TNF-α, IL-1, IL-6, and/or IL-8 aretransmitted to a controller associated with the extracorporeal device.The controller compares the current levels of TNF-α, IL-1, IL-6, and/orIL-8 with one or more target values, e.g., desired concentrations ofTNF-α, IL-1, IL-6, and/or IL-8. As appropriate, the controller divertsall or part of the whole blood or plasma to one or more treatmentchambers in the extracorporeal device to treat the blood to achieve atarget value of the one or more inflammatory mediators. Diversion ofblood into one or more treatment chambers of the extracorporeal devicecontinues until the target value of the one or more inflammatorymediators has been achieved.

The one or more treatment chambers include one or more binding agentsconfigured to bind and sequester one or more inflammatory mediators. Inthis example, the binding agents are polymer based, artificial bindingsubstrates formed by molecular imprinting and are designed toselectively bind and sequester TNF-α, IL-1, IL-6, and/or IL-8 within theone or more treatment chambers. In one configuration, all or part of thewhole blood or plasma passes through a single treatment chamber thatincludes binding agents directed towards multiple inflammatorymediators. In an alternative configuration, all or part of the wholeblood or plasma passes through multiple treatment chambers, eachconfigured with a binding agent directed to one specific inflammatorymediator. For example, all or part of the whole blood may passsequentially through a series of treatment chambers configured toselectively bind and sequester TNF-α, IL-1, IL-6, or IL-8, respectively,to achieve a target value for each of these inflammatory mediators. Thetreated blood is returned to the peripheral circulation of the subjectthrough a conduit leaving the extracorporeal device.

Example 6

Extracorporeal Device for Sensing, Binding and Altering InflammatoryMediators

A method for treating an inflammatory condition or disease is providedand includes an extracorporeal device configured to sense one or moreinflammatory mediators, and to bind and alter the functional structureof the one or more inflammatory mediators in the peripheral blood of asubject to achieve a desired target value. The extracorporeal deviceincludes a flow route for extracting blood from a subject, a componentfor optionally fractionating the whole blood into separate components,one or more sensors for sensing the levels of one or more inflammatorymediators, a controller to receive data regarding the sensed levels ofone or more inflammatory mediators, one or more binding elementsincorporated into one or more treatment chambers, one or more reactivecomponents designed to alter the functional structure of one or moreinflammatory mediators, and a flow route for returning the processedblood to the subject.

Blood is removed from a subject through a catheter inserted into thesubject. The catheter is further attached to one or more conduits thatflow into the extracorporeal device. The whole blood is fractionated bycentrifugation and/or filtration within the extracorporeal bloodprocessing device to isolate plasma. The plasma containing inflammatorymediators but free of blood cells is optionally further fractionatedusing one or more semi-permeable membranes to isolate a plasma fractionenriched in blood components ranging in molecular weight from about1,000 to about 150,000 dalton.

The fractionated plasma is passed over one or more sensors associatedwith the extracorporeal device. The one or more sensors include one ormore recognition elements and are configured to sense one or moreinflammatory mediators. In this example, the recognition elements linkedto the one or more sensors are antibodies configured to selectivelyrecognize the proinflammatory mediators INF-α, IL-1, IL-6, and IL-8. Thelevels of TNF-α, IL-1, IL-6, and/or IL-8 in the plasma are sensed, andthe data are transmitted to a controller associated with theextracorporeal device. The controller compares the current levels ofTNF-α, IL-1, IL-6, and/or IL-8 with one or more target values, e.g.,desired concentrations of TNF-α, IL-1, IL-6, and/or IL-8. Asappropriate, the controller diverts all or part of the fractionatedplasma to one or more treatment chambers in the extracorporeal device totreat the blood to achieve a target value of the one or moreinflammatory mediators.

The one or more treatment chambers include one or more binding elementsthat capture and retain one or more inflammatory mediators in thetreatment chamber. In this example, the binding agents areoligonucleotide-based DNA aptamers directed towards binding theinflammatory mediators TNF-α, IL-1, IL-6, and IL-8. The aptamersdirected towards TNF-α, IL-1, IL-6, and IL-8 are attached to beads usingone or more of the crosslinking methods described herein. Theaptamer-modified beads are retained within the treatment chamber of theextracorporeal device via size exclusion using a membrane or screen thatis otherwise permeable to blood components. The pro-inflammatorymediators TNF-α, IL-1, IL-6, and IL-8 in the plasma bind to theaptamer-modified beads and are retained in the treatment chamber.Non-bound components of the plasma flow out of the treatment chamber andare returned to the peripheral blood of the subject.

Altering the functional structure of TNF-α, IL-1, IL-6, and/or IL-8occurs in the treatment chamber by denaturation and/or degradation ofthe pro-inflammatory mediators using one or more proteases configured tocleave one or more peptide bonds of the bound TNF, IL-1, IL-6 and/orIL-8. The treatment chamber is isolated from the peripheral blood flowof the subject and flooded with the one or more proteases to digest thepeptide bonds of the one or more inflammatory mediators. Followingprotease treatment, the treatment chamber may be washed with anappropriate physiological solution, e.g. 0.9% saline solution, prior toreceiving more plasma for processing.

The fractionated plasma in which the one or more inflammatory mediatorshas been altered or destroyed is combined with other components of thewhole blood and may be returned to the subject.

Example 7

Extracorporeal Device for Sensing Inflammatory Mediators andControllably Releasing Inflammatory Modulators

A method for treating an inflammatory condition or disease is providedand includes an extracorporeal device designed to sense one or moreinflammatory mediators in the peripheral blood of a subject and tocontrollably release one or more inflammatory modulators to achieve adesired target value of the one or more inflammatory mediators. Theextracorporeal device includes one or more sensors for sensing one ormore inflammatory mediators, a controller to receive data regarding thesensed levels of the one or more inflammatory mediators, one or morereservoirs containing one or more inflammatory mediators controllablyopened or closed by the controller, and optionally a transmitter forwirelessly transmitting the sensed levels of one or more inflammatorymediators to the subject and/or one or more caregivers.

Blood is removed from a subject through a catheter inserted into thesubject. The catheter is further attached to one or more conduits thatflow into the extracorporeal device. All or part of the flow of wholeblood, plasma, or fractionated plasma, may be analyzed for the level ofone or more inflammatory mediators. In some aspects, the extracorporealdevice includes a component for separating a small sample of blood fromthe bulk blood flow. The small stream of blood passes through acompartment of the device that includes one or more sensors for sensingthe level of one or more inflammatory mediators. The level of one ormore inflammatory mediators in a small sample of blood flow is believedto be representative of the level of one or more inflammatory mediatorsin the blood as a whole.

The one or more sensors associated with the device may use any of anumber of methodologies including surface plasmon resonance,piezoelectric, and/or molecular beacons as described herein. In oneconfiguration, the one or more sensors are an array of sensor chipscapable of detecting multiple inflammatory mediators simultaneously. Thesensor chips include recognition elements that interact with any of anumber of proinflammatory mediators, e.g, TNF-α, IL-1, IL-6, IL-8, IL-12and any of a number of anti-inflammatory mediators, e.g., IL-4, IL-10,IL-13 and TGFβ. In this example, the recognition elements are antibodiesselective for a given inflammatory mediator, but could also includeaptamers, receptors, ligands or any other biomolecule that binds to aninflammatory mediator. As peripheral blood from the subject passes overthe sensor surface, inflammatory mediators present in the blood interactwith the cognate recognition elements on the sensor chips, causing asignal to be transmitted to the controller. The controller compares thedata regarding the current levels of one or more inflammatory mediatorsin the peripheral blood of a subject with target values, e.g., desiredconcentrations for the one or more inflammatory mediators. Asappropriate, the controller signals release of one or more reactivecomponents, e.g., one or more inflammatory modulators, from one or morereservoirs associated with the extracorporeal device to achieve thetarget values. Alternatively, the controller may transmit a message tothe subject and/or a third party individual regarding the level of oneor more inflammatory mediators and the assessment of which reactivecomponents or modulators, if any, should be released into the peripheralblood of the subject to modulate an inflammatory response. In response,the subject and/or a third party individual may choose to administer oneor more modulators into the peripheral blood of the subject.

The intracorporeal device includes one or more reservoirs containing oneor more inflammatory modulators. In this example, the inflammatorymodulators alter the functional levels of any of a number ofproinflammatory mediators, e.g, TNF-α, IL-1, IL-6, IL-8, IL-12 and anyof a number of anti-inflammatory mediators, e.g., IL-4, IL-10, IL-13 andTGF-β and are contained in separate reservoirs within the extracorporealdevice. For example, one set of reservoirs contains a soluble TNF-αreceptor, e.g., etanercept; a second set of reservoirs contains an IL-1receptor antagonist, e.g., anakinra; a third set of reservoirs containsrecombinant IL-10; and a fourth set of reservoirs contains recombinantTGF-β. Release of the inflammatory modulators from the one or morereservoirs is controlled by signals sent from the controller and isbased on the sensed levels of TNF-α, IL-1, IL-10 and/or TGF-β in theperipheral blood. In this configuration, the levels of theproinflammatory mediators TNF-α and IL-1 may be controllably decreasedand the levels of the anti-inflammatory mediator IL-10 and TGF-β may becontrollably increased to achieve appropriate target values of eachinflammatory mediator.

Example 8

Combined Intracorporeal and Extracorporeal Device for Sensing, Bindingand Inactivating Inflammatory Mediators

A method for treating an inflammatory condition or disease is providedand includes a device having combined intracorporeal and extracorporealcomponents to sense one or more inflammatory mediators, and to bind andalter the one or more inflammatory mediators in the peripheral blood ofa subject to achieve a target value. The device includes one or moreintracorporeal components to sense the level one or more inflammatorymediators in the peripheral blood, a controller to receive dataregarding the sensed levels of the one or more inflammatory mediators,an extracorporeal component to withdraw the peripheral blood, one ormore reactive components to alter the functional structure of the one ormore inflammatory mediators, and a transmitter for wirelesslytransmitting the sensed levels of one or more inflammatory mediators tothe subject and/or one or more caregivers.

The one or more intracorporeal components of the device are placed in orproximal to one or more blood vessels of a subject. The intracorporealcomponent of the device may be a stent-like structure that resides in afixed position in a blood vessel. Alternatively, the intracorporealcomponent of the device may be free to travel in the lumen of one ormore blood vessels. See, e.g., U.S. Patent Application 2007/0156211 A1,which is incorporated herein by reference.

The one or more intracorporeal components of the device include one ormore sensors for sensing the level of the one or more inflammatorymediators in the peripheral blood of a subject. The sensors can be anyof a number of sensor types as described herein. The one or more sensorsinclude one or more recognition elements that interact with one or moreinflammatory mediators as described herein. The sensed data aretransmitted wirelessly to a controller associated with theextracorporeal component of the device.

The controller receives the data regarding the level of one or moreinflammatory mediators in the peripheral blood of a subject. Thecontroller compares the current levels of one or more inflammatorymediators with a target value, e.g., a desired concentration orconcentration range for one or more inflammatory mediators. Based on thecomparison between the current levels and the target value, thecontroller automatically initiates extracorporeal treatment of the bloodto achieve the appropriate target values of the one or more inflammatorymediators to control an inflammatory response in a subject. In thisexample, the subject is constantly connected to the extracorporealcomponent of the device via a flow route or conduit and the controlleris able to automatically initiate withdrawal of blood from the subjectand into the extracorporeal treatment chamber. Alternatively, thecontroller sends data regarding the current levels of one or moreinflammatory mediators and the desired target values to the subjectand/or a caregiver. The subject and/or caregiver may choose to initiateextracorporeal treatment of the blood based on the information providedby the controller.

The extracorporeal component of the device releases or activates any ofa number of reactive components designed to alter the functionalstructure of one or more inflammatory mediators as described herein. Thereactive component may be non-selective, e.g., a filtering system thatnon-selectively captures and sequesters one or more inflammatorymediators. Alternatively, the reactive component may be binding agentssuch as antibodies or aptamers that bind specific inflammatorymediators, e.g., TNF-α and IL-1 and retain them in the treatmentchamber. Other reactive components, e.g., one or more of a denaturingagent, a degradative agent, an enzyme, a chemical, an energy source, amodulator, or a combination thereof may be used to alter the functionalstructure of the inflammatory mediators retained in the treatmentchamber. The treated blood having attained a target level of the one ormore inflammatory mediators is returned to the peripheral circulation ofthe subject through a conduit leaving the extracorporeal device.

While various aspects and embodiments have been disclosed herein, otheraspects and embodiments will be apparent to those skilled in the art.The various aspects and embodiments disclosed herein are for purposes ofillustration and are not intended to be limiting, with the true scopeand spirit being indicated by the following claims.

1. A method for treating an inflammatory disease or condition in asubject comprising: providing a device configured to communicate with atleast a portion of a peripheral blood of the subject, the deviceincluding a sensor configured to detect one or more inflammatorymediators in peripheral blood of a subject; a controller incommunication with the sensor, the controller configured to controlinteraction between one or more reactive components and the one or moreinflammatory mediators, the one or more reactive components configuredto alter a functional structure of the one or more inflammatorymediators in the peripheral blood, wherein the controller is configuredto control the interaction between the one or more reactive componentsand the one or more inflammatory mediators to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject, and the controller is in communication with a reservoirconfigured to release the one or more reactive components into theperipheral blood, and flowing blood through the device to treat theinflammatory disease or condition in the subject.
 2. The method of claim1, further including a treatment chamber configured to receive at leasta portion of the peripheral blood through a flow route, the controllerconfigured to control flow of peripheral blood through the flow routeinto the treatment chamber, and the treatment chamber including one ormore reactive components configured to alter a functional structure ofthe one or more inflammatory mediators in the peripheral blood.
 3. Themethod of claim 2, wherein the controller is configured to controlaccess to the treatment chamber by the peripheral blood.
 4. The methodof claim 2, wherein the one or more reactive components include one ormore of a denaturing agent, a degradative agent, binding agent, or anenergy source.
 5. The method of claim 4, wherein the one or more bindingagents on a matrix adapted to the treatment chamber are configured tosequester at least one of the one or more inflammatory mediators fromthe blood.
 6. The method of claim 5, wherein the matrix includes one ormore of a specific binding ligand or a hydrophobic surface.
 7. Themethod of claim 4, wherein the one or more reactive components increasesan activity of an intermediate which decreases the activity of one ormore of TNF-α, IL-1, IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, IFN-γ, LIF,MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin, or endotoxin.
 8. The method ofclaim 4, wherein the energy source includes at least one of microwaveirradiation, gamma irradiation, electron beam irradiation,vibrational/frequency irradiation, or atmospheric pressure glowdischarge.
 9. The method of claim 8, wherein the vibrational/frequencyirradiation includes a set of differing energy inputs specificallydirected to the one or more inflammatory mediators, wherein the set ofdiffering energy inputs selectively resonates a plurality of resonantstructures in the one or more inflammatory mediators, and wherein theresonance controllably alters or reduces the activity of the one or moreinflammatory mediators in the peripheral blood of the subject.
 10. Themethod of claim 9, wherein the one or more inflammatory mediators aremodified with a functional group configured to be responsive to the setof differing energy inputs.
 11. The method of claim 4, wherein the oneor more degradative agents include at least one of an enzyme, coenzyme,enzyme complex, catalytic antibody, proteasome, strong acid, strongbase, radical, photoactivatable agent, drug, cell, or radical ion. 12.The method of claim 4, wherein the one or more reactive componentsinclude at least one modulator of the one or more inflammatory mediatorsconfigured to alter the functional structure of the one or moreinflammatory mediators in the peripheral blood of the subject.
 13. Themethod of claim 1, wherein the one or more reactive components includeone or more of a denaturing agent, a degradative agent, a binding agent,or an energy source.
 14. The method of claim 13, wherein the one or morebinding agents include one or more of antibodies, receptors, or cognatesconfigured to bind to at least one of the one or more inflammatorymediators.
 15. The method of claim 13, wherein the one or more bindingagents include one or more of lectin, binding protein, catalyticantibody, catalytic aptamer, protease conjugate, or photoactivatableagent conjugate.
 16. The method of claim 13, wherein the one or morereactive components increases an activity of an intermediate whichdecreases the activity of one or more of TNF-α, IL-1, IL-1β, IL-6, IL-8,IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin,or endotoxin.
 17. The method of claim 13, wherein the one or morereactive components decreases an activity of an intermediate whichdecreases the activity of one or more of TNF-α, IL-1, IL-1β, IL-6, IL-8,IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a , C5-a, exotoxin,or endotoxin.
 18. The method of claim 13, wherein the one or morereactive components is configured to modulate an activity of one or moreof anaphylatoxins, cytokines, chemokines, leukotrienes, prostaglandins,complement, coagulation factors, or proinflammatory cytokines.
 19. Themethod of claim 13, wherein the one or more degradative agents includeat least one of an enzyme, coenzyme, enzyme complex, catalytic antibody,proteasome, strong acid, strong base, radical, photoactivatable agent,drug, cell, or radical ion.
 20. The method of claim 13, wherein the oneor more reactive components include at least one modulator of the one ormore inflammatory mediators configured to alter the functional structureof the one or more inflammatory mediators in the peripheral blood of thesubject.
 21. The method of claim 1, wherein the sensor and thecontroller are configured to control levels of the one or moreinflammatory mediators to substantially attain the target value.
 22. Themethod of claim 1, wherein the sensor includes a biosensor, chemicalsensor, physical sensor, or optical sensor.
 23. The method of claim 1,wherein the sensor is configured to target the device to a site havingan elevated level of the inflammatory mediators.
 24. The method of claim1, wherein the sensor is configured to detect one or more ofanaphylatoxin, cytokine, chemokine, leukotriene, prostaglandin,complement, coagulation factor, or proinflammatory cytokine.
 25. Themethod of claim 1, wherein the one or more reactive components isconfigured to alter a functional structure of the one or moreinflammatory mediators in the peripheral blood, and wherein the one ormore reactive components is configured to decrease an activity of one ormore of TNF-α, IL-1, IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, IFN-γ, LIF,MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin, or endotoxin.
 26. The method ofclaim 1, wherein the one or more reactive components configured to altera functional structure of the one or more inflammatory mediators in theperipheral blood includes acoustic energy or electronic energy.
 27. Themethod of claim 26, wherein the energy source includes ultrasound. 28.The method of claim 1, wherein the device is extracorporeal.
 29. Amethod for treating an inflammatory disease or condition in a subjectcomprising: providing a device configured to communicate with at least aportion of a peripheral blood of the subject, the device including asensor configured to detect one or more inflammatory mediators inperipheral blood of a subject; a controller in communication with thesensor, a means for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject, and flowing blood through the device to treat theinflammatory disease or condition in the subject, wherein the targetvalue includes a desired ratio of concentrations of two or moreinflammatory mediators in the peripheral blood.
 30. A method fortreating an inflammatory disease or condition in a subject comprising:providing a device configured to communicate with at least a portion ofa peripheral blood of the subject, the device including a sensorconfigured to detect one or more inflammatory mediators in peripheralblood of a subject; a controller in communication with the sensor, ameans for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject, and flowing blood through the device to treat theinflammatory disease or condition in the subject, wherein the sensor andthe controller are configured to control levels of the one or moreinflammatory mediators to limit a deviation from the target value. 31.The method of claim 30, wherein the deviation is determined by aweighted least squares fit.
 32. A method for treating an inflammatorydisease or condition in a subject comprising: providing a deviceconfigured to communicate with at least a portion of a peripheral bloodof the subject, the device including a sensor configured to detect oneor more inflammatory mediators in peripheral blood of a subject; acontroller in communication with the sensor, a means for modulating aphysiological effect of the one or more inflammatory mediatorsresponsive to the controller, the controller configured to adjust themodulating means to achieve a target value of the detected one or moreinflammatory mediators in the peripheral blood of the subject, andflowing blood through the device to treat the inflammatory disease orcondition in the subject, wherein the sensor includes a physical sensorincluding one or more of an aptamer, antibody, or receptor.
 33. A methodfor treating an inflammatory disease or condition in a subjectcomprising: providing a device configured to communicate with at least aportion of a peripheral blood of the subject, the device including asensor configured to detect one or more inflammatory mediators inperipheral blood of a subject; a controller in communication with thesensor, a means for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject, and flowing blood through the device to treat theinflammatory disease or condition in the subject, wherein the sensorincludes a physical sensor including one or more of a recognition-basedsubstrate, an aptamer-based substrate, an antibody-based substrate,surface plasmon resonance, genetically-modified cells, orgenetically-modified cells with receptor-linked signaling.
 34. Themethod of claim 33, wherein the genetically-modified cells includereceptor-linked signaling by fluorogen-activating proteins.
 35. A methodfor treating an inflammatory disease or condition in a subjectcomprising: providing a device configured to communicate with at least aportion of a peripheral blood of the subject, the device including asensor configured to detect one or more inflammatory mediators inperipheral blood of a subject; a controller in communication with thesensor, a means for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject, and flowing blood through the device to treat theinflammatory disease or condition in the subject, wherein the sensor isconfigured to detect cytokines, T-lymphocytes, B-lymphocytes, orantibodies.
 36. A method for treating an inflammatory disease orcondition in a subject comprising: providing a device configured tocommunicate with at least a portion of a peripheral blood of thesubject, the device including a sensor configured to detect one or moreinflammatory mediators in peripheral blood of a subject; a controller incommunication with the sensor, a means for modulating a physiologicaleffect of the one or more inflammatory mediators responsive to thecontroller, the controller configured to adjust the modulating means toachieve a target value of the detected one or more inflammatorymediators in the peripheral blood of the subject, and flowing bloodthrough the device to treat the inflammatory disease or condition in thesubject, wherein the sensor is further configured to detect bodytemperature, vital signs, edema, oxygen level, or pathogen/toxin levelof the subject.
 37. A method for treating an inflammatory disease orcondition in a subject comprising: providing a device configured tocommunicate with at least a portion of a peripheral blood of thesubject, the device including a sensor configured to detect one or moreinflammatory mediators in peripheral blood of a subject; a controller incommunication with the sensor, a means for modulating a physiologicaleffect of the one or more inflammatory mediators responsive to thecontroller, the controller configured to adjust the modulating means toachieve a target value of the detected one or more inflammatorymediators in the peripheral blood of the subject, and flowing bloodthrough the device to treat the inflammatory disease or condition in thesubject, wherein the one or more reactive components include one or morebinding agents on a matrix adapted to the treatment chamber areconfigured to sequester at least one of the one or more inflammatorymediators from the blood and the one or more binding agents include oneor more of antibodies, receptors, or cognates configured to bind to atleast one of the one or more inflammatory mediators.
 38. A method fortreating an inflammatory disease or condition in a subject comprising:providing a device configured to communicate with at least a portion ofa peripheral blood of the subject, the device including a sensorconfigured to detect one or more inflammatory mediators in peripheralblood of a subject; a controller in communication with the sensor, ameans for modulating a physiological effect of the one or moreinflammatory mediators responsive to the controller, the controllerconfigured to adjust the modulating means to achieve a target value ofthe detected one or more inflammatory mediators in the peripheral bloodof the subject, and flowing blood through the device to treat theinflammatory disease or condition in the subject, wherein the one ormore reactive components include one or more binding agents on a matrixadapted to the treatment chamber are configured to sequester at leastone of the one or more inflammatory mediators from the blood and the oneor more binding agent includes at least one of lectin, binding protein,catalytic antibody, catalytic aptamer, protease conjugate, orphotoactivatable agent conjugate.
 39. A method for treating aninflammatory disease or condition in a subject comprising: providing adevice configured to communicate with at least a portion of a peripheralblood of the subject, the device including a sensor configured to detectone or more inflammatory mediators in peripheral blood of a subject; acontroller in communication with the sensor, a means for modulating aphysiological effect of the one or more inflammatory mediatorsresponsive to the controller, the controller configured to adjust themodulating means to achieve a target value of the detected one or moreinflammatory mediators in the peripheral blood of the subject, andflowing blood through the device to treat the inflammatory disease orcondition in the subject, wherein the one or more reactive componentsinclude one or more binding agents on a matrix adapted to the treatmentchamber configured to sequester at least one of the one or moreinflammatory mediators from the blood, and the matrix includes one ormore of beads, cells, vesicles, filters, hydrogel polymers,microparticles, nanoparticles, adsorbent, or synthetic polymers.
 40. Themethod of claim 39, wherein the specific binding ligand or thehydrophobic surface includes one or more of nucleic acid aptamers,peptide aptamers, molecular imprinting polymer, antibodies or fragmentsthereof, high affinity mimetics, synthetic binding molecules, orreceptor binding molecules.
 41. A method for treating an inflammatorydisease or condition in a subject comprising: providing a deviceconfigured to communicate with at least a portion of a peripheral bloodof the subject, the device including a sensor configured to detect oneor more inflammatory mediators in peripheral blood of a subject; acontroller in communication with the sensor, a means for modulating aphysiological effect of the one or more inflammatory mediatorsresponsive to the controller, the controller configured to adjust themodulating means to achieve a target value of the detected one or moreinflammatory mediators in the peripheral blood of the subject, andflowing blood through the device to treat the inflammatory disease orcondition in the subject, wherein the one or more reactive componentsinclude one or more binding agents on a matrix adapted to the treatmentchamber are configured to sequester at least one of the one or moreinflammatory mediators from the blood and the matrix includes one ormore of a lectin, binding protein, receptor, antibody, catalyticantibody, catalytic aptamer, protease conjugate, or photoactivatableagent conjugate.
 42. A method for treating an inflammatory disease orcondition in a subject comprising: providing a device configured tocommunicate with at least a portion of a peripheral blood of thesubject, the device including a sensor configured to detect one or moreinflammatory mediators in peripheral blood of a subject; a controller incommunication with the sensor, a means for modulating a physiologicaleffect of the one or more inflammatory mediators responsive to thecontroller, the controller configured to adjust the modulating means toachieve a target value of the detected one or more inflammatorymediators in the peripheral blood of the subject, and flowing bloodthrough the device to treat the inflammatory disease or condition in thesubject, wherein the one or more reactive components include one or morebinding agents that decrease an activity of an intermediate thatdecrease the activity of one or more of TNF-α, IL-1, IL-1β, IL-6, IL-8,IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1, C3-a, C5-a, exotoxin,or endotoxin.
 43. A method for treating an inflammatory disease orcondition in a subject comprising: providing a device configured tocommunicate with at least a portion of a peripheral blood of thesubject, the device including a sensor configured to detect one or moreinflammatory mediators in peripheral blood of a subject; a controller incommunication with the sensor, a means for modulating a physiologicaleffect of the one or more inflammatory mediators responsive to thecontroller, the controller configured to adjust the modulating means toachieve a target value of the detected one or more inflammatorymediators in the peripheral blood of the subject, and flowing bloodthrough the device to treat the inflammatory disease or condition in thesubject, wherein the means for modulating a physiological effect of theone or more inflammatory mediators includes one or more reactivecomponents configured to alter a functional structure of the one or moreinflammatory mediators in the peripheral blood wherein the one or morereactive components is configured to modulate an activity of one or moreof anaphylatoxins, cytokines, chemokines, leukotrienes, prostaglandins,complement, coagulation factors, or proinflammatory cytokines.
 44. Amethod for treating an inflammatory disease or condition in a subjectcomprising: providing a device configured to communicate with at least aportion of a peripheral blood of the subject, the device including asensor configured to detect one or more inflammatory mediators inperipheral blood of a subject: a controller in communication with thesensor, a treatment chamber configured to receive at least a portion ofthe peripheral blood through a flow route, the controller configured tocontrol flow of peripheral blood through the flow route into thetreatment chamber, and the treatment chamber including one or morereactive components configured to alter a functional structure of theone or more inflammatory mediators in the peripheral blood, thecontroller configured to adjust flow to the treatment chamber to achievea target value of the detected one or more inflammatory mediators in theperipheral blood of the subject, and flowing blood through the device totreat the inflammatory disease or condition in the subject, and whereinthe treatment chamber includes a source for producing the one or morereactive components.
 45. The method of claim 44, wherein the sourceincludes at least one reservoir and at least one producer.
 46. Themethod of claim 45, wherein the at least one producer includes at leastone encapsulated cell.
 47. The method of claim 46, wherein the at leastone encapsulated cell produces the one or more reactive components. 48.A method for treating an inflammatory disease or condition in a subjectcomprising: providing a device configured to communicate with at least aportion of a peripheral blood of the subject, the device including asensor configured to detect one or more inflammatory mediators inperipheral blood of a subject; a controller in communication with thesensor, a treatment chamber configured to receive at least a portion ofthe peripheral blood through a flow route, the controller configured tocontrol flow of peripheral blood through the flow route into thetreatment chamber, the treatment chamber including one or more reactivecomponents configured to alter a functional structure of the one or moreinflammatory mediators in the peripheral blood, the controllerconfigured to adjust flow to the treatment chamber to achieve a targetvalue of the detected one or more inflammatory mediators in theperipheral blood of the subject, and flowing blood through the device totreat the inflammatory disease or condition in the subject, and whereinthe treatment chamber includes one or more reservoirs including one ormore reactive components.